1,302 Matching Annotations
  1. Last 7 days
    1. Characterization of embelin isolated from E.ribes
    2. . Extraction and isolation of embelin from E. ribes
    3. High Performance Liquid Chromatography (HPLC) analysis
    4. High Performance Thin Layer Chromatography (HPTLC) analysis
    5. . Thin Layer Chromatography (TLC) analysis
    6. Preliminary phytochemical screening (Ali, 1998; Evans, 2002)
    7. Microbial Contamina
    8. Foaming Index
    9. pH values
    10. Ash values
    11. Extractive value
    12. . Physico-chemical standardization
    13. Standardization of ethanolic extract of E.ribes (WHO, 1998; IndianPharmacopoeia, 1996)
    14. Preparation of ethanolic extract of E.ribes
    15. PLANT MATERIAL
    1. Estimation of Stevioside
    2. Extraction from the plant material
    3. Extraction from the plant material
    4. Extraction from the plant material
    5. Qmmtification of stevioside
    6. Estimation of Steviol
    7. Extraction from plant material
    8. Quantification of steviol
    9. Estimation of free aminoacids
    10. Estimation of soluble proteins
    11. Estimation of sugars
    12. Estimation of total phenols
    13. Determination of moisture
    14. Analytical methods
    15. The experimental part of the study was categorized into five sections for the convenience of reference viz. analytical, toxicological, molecular, biochemical and genomic quantitation.
    16. Plant material
    17. e5 tevia-the natural smeetenei
    1. Per-cent Overlap
    2. Compogram
    3. F-and t-Ratios
    4. Chi-Square
    5. Somatotype Categories
    6. Comparative Statistics
    7. Somatotype Attitudinal Mean (SAM)
    8. Somatotype Attitudinal Distance (SAD)
    9. Somatotype Dispersion Mean (SDM)
    10. Somatotype Dispersion Distances (SDD)
    11. Somatoplot Coordinates
    12. Mean Somatotypes (5)
    13. Descriptive Statistics
    14. Statistical Analysis
    15. Central:
    16. Mesomorphic ectomorph:
    17. Endomorphic ectomorph:
    18. Ectomorphic mesomo~ph:
    19. Endomorphic mesomorph:
    20. Ectomorphic endomorph:
    21. Mesomorphic endomor~:
    22. Endomorph-ectom9££E:
    23. Mesomorph-endomorph:
    24. Mesomorph-endomorph:
    25. Balanced ectomorph:
    26. Balanced mesomorph:
    27. Balanced endomorph:
    28. SOHATOCHART
    29. Somatotyping Children
    30. Limitations of the Rating Form
    31. Third Component Rating
    32. Second Component Rating
    33. First Component Rating
    34. SOMATOTYPING
    35. Calf
    36. Biceps
    37. Femur
    38. Humerus
    39. Calf
  2. Jun 2019
    1. Primary culture of E. coli was grown in LB medium containing either ampicillin (Amp) and/or kanamycin (Kan) to final concentration of 100 j..tg/ml and 25 J..tg/ml respectively. Depending on the vector construct, antibiotics were used for expression of different proteins as described in Table 3.1. Medium was inoculated with 1 ml glycerol stock of E. coli and incubated overnight at 37 oc at 200 rpm.
    2. Preparation of primary culture of E. coli cells
    1. Blood was drawn from appropriate source into heparinised tubes. The blood sample was centrifuged at 4000 rpm for 15 min ( 4 °C). The supernatant was discarded, and the erythrocytes (pellet) were subsequently washed thrice with chilled isotonic buffer [0.01 M PBS (pH 7.4)] by centrifugation at 4000 rpm and 4°C for 15 min. The washed erythrocytes were lysed in water. The resultant red cell lysate was then dialyzed extensively against PBS (pH 7.4) at 4°C to obtain stripped hemoglobin (hemoglobin devoid of bound allosteric modulators like BPG). The stripped hemoglobin was then loaded onto a pre-equilibrated DE52 column (30cm x 15cm) after extensive dialysis against 0.05 M tris acetate buffer (pH 8.5). The protein was eluted from the column employing a linear gradient of 500 ml each of 0.05 M tris acetate (pH 8.5) and 0.05 M tris acetate (pH 7) at a flow rate of 50 ml/hour. The purified hemoglobin was estimated spectrophotometrically at 540 nm (molar extinction coefficient= 53236 cm-1/M) and stored at -70°C till further use.
    2. Purification of hemoglobin from bloo
  3. May 2019
    1. Vancouver-based Contextual Genomics has launched two molecular hotspot assays for detecting genomic mutations in blood and solid tumors. <!--//--><![CDATA[// ><!-- document.addEventListener("googletagEvent", function () { let hideOnMobile = "0"; let isMobileQuery = false; if (typeof window.dataLayerValues === "undefined" || !window.dataLayerValues.hasOwnProperty('isMobileQuery') || window.dataLayerValues.isMobileQuery) { isMobileQuery = window.dataLayerValues.isMobileQuery; } // Don't display the ad unit if this is a mobile browser and we're supposed to hide the unit in mobile view. if (isMobileQuery && hideOnMobile === '1') { return; } let hideOnDesktop = "0"; // Don't display this ad unit if this is a desktop broser and we're supposed to hide the mobile ad unit if(!isMobileQuery && hideOnDesktop === '1') { return; } googletag.cmd.push(function () { googletag.display('content-embed-one'); }); }); //--><!]]> The new version of the company’s Find It solid tumor panel now screens for 146 somatic genome alterations, and 23 exons in 30 cancer-associated genes, to help identify precision cancer treatments and recognize drug-resistant mutations. New additions to the panel include tests for mutations in the POLE gene, which have been associated with colorectal cancer as well as immunodeficiency.

      31-05-2019

    1. After breakfast they made ready to say farewell, as nearly heavy of heart3 as was possible on such a morning; cool, bright, and clean under a washed autumn sky of thin blue. The air came fresh from the North-West.They rode off along a path and looked out from the hill-top over lands under the morning. It was now as clear and far-seen as it had been veiled and misty when they stood upon the knoll in the Forest. They took a deep draught of the air.4Their way wound along the floor of the hollow, and round the green feet of a steep hill into another deeper and broader valley. As they journeyed the sun mounted, and grew hot. Each time they climbed a ridge the breeze seemed to have grown less. When they caught a glimpse of the country westward the distant Forest seemed to be smoking, as if the fallen rain was steaming up again. A shadow now lay round the edge of sight, a dark haze above which the sky was like a blue cap.5 On that side the hills were higher and looked down upon them; and all those hills were crowned with green mounds, and on some were standing stones, pointing upwards like jagged teeth out of green gums. The view was somehow disquieting; so they turned from the sight and went down into the hollow circle. In the midst of it there stood a single stone, standing tall under the sun above, and at this hour casting no shadow. They set their backs6 against the east side of the stone. It was cool, as if the sun had had no power to warm it. There they took food and drink.Riding over the hills, and eating their fill,7 lying a little too long; these things are, perhaps, enough to explain what happened. How­ever, that may be: they woke suddenly from a sleep they had never meant to take. The standing stone was cold, and it cast a long pale shadow. The sun was gleaming through the mist; north, south, and east, the fog was thick, cold and white. The air was silent, heavy and chill.
    1. Humanities faculty, unlike their STEM counterparts, do not have labs. We do not have a place for our work and no one sees our process.

      Is this implying that people do see progress in labs? Or that somehow labs are in a way accessible for people to come in and view academic research in progress? If that's a thing that happens, I'd love to check in on the labs of more advanced students, but I have a strong feeling that simply asking to be in a lab and watch people work will be met with quite a bit of resistance.

    2. However, that work (and it is intellectual labor) is invisible and largely undervalued.

      In my microbiology lab in the January semester I realized for the first time exactly how much work goes into a paper. It gave me a healthy respect for published academics, as well as made me realize, immediately, I do not want to stay in academia my whole life. Some people are incredible with the amount of effort they put into their research.

    3. Academics are constantly being told that they need to make their work more relevant and accessible to the public. Blogging about your work hits both of those marks. It also means that you have to translate your work from academese to language that non-academics will understand (i.e. jargon) and also foreground the relevance of your work. You have to tell people why your work is important and what it adds to the world.

      Do you ever wish you read the whole article before annotating because you read one paragraph down and find out the article says the exact thing you said in your annotation? Yeah. Well, at least I feel validated in my constant search for accessible academic content.

    4. As a result, I suggest that one thing that all humanities scholars can do to take a baby step in the direction of digital humanities is to maintain a blog about their research.

      Oh, what a coincidence that our major project in this class is to maintain a blog about our findings! Joking aside, I wish articles came with a link to a blog about the research involved in them. No matter how many times I read and re-read the methods section I never can seem to fully understand what the researchers were doing, because I'm an undergraduate just scratching the surface of topics. A blog would have more casual details and wouldn't assume the audience knows a lot already and would allow me to learn without having to delve for 80 hours down a rabbit hole about a specific enzyme in one microbe to figure out why it was even mentioned. Or maybe I'm just not a natural born student!

    1. “People will use this data in ways we can’t even imagine yet,” Mr. Stowell said, “and I think that is one of the most exciting developments in the humanities.”

      I keep coming back to history in my annotations, and honestly the article could work as a reading in a history class too. This kind of collection of data; of sources for the future could do wonders for future historians. Digital records, especially those online, don't burn or get water damaged or get eaten by moths. I think it's very important that we consider our digital footprints in a historical sense, from our own personal data (which I can see functioning much the same way as diaries do for historians now) to larger projects such as the tapestry mentioned above.

    2. Mr. Edelstein said that many of his senior colleagues view his work as whimsical, the result of playing with technological toys. But he argues such play can lead to discoveries.

      As he should; he's correct. Technological advances come from "playing with technological toys" all the time, it's no stretch of the imagination to assume academic advances would as well. Of course people are always resistant to change; it's in our nature, but using the tools at our disposal to improve our work is a part of academia.

    3. “You would think if England was this fountainhead of freedom and religious tolerance,” he said, “there would have been greater continuing interest there than what our correspondence map shows us.”

      While I am not surprised that the extent of England's greatness was greatly exaggerated (given our colonial, euro, and white -centric views of history) it's very important to have the data and evidence to back it up.

    4. Even historians, who have used databases before, have been slow to embrace the trend. Just one of the nearly 300 main panels scheduled for next year’s annual meeting of the American Historical Association covers digital matters.

      We explored the expansion of digital records briefly in HIST211 in the January semester. One of the issues with history is having very few (if any) primary sources, but a bigger issue is that they often contradict each other. Digital databanks and scans of old documents and even sites like ancestry.ca have broadened sources available to historians but they also cause more contradictions to be found, making the reconstruction of any historical event/period potentially more difficult.

    5. Mr. Bobley said the emerging field of digital humanities is probably best understood as an umbrella term covering a wide range of activities, from online preservation and digital mapping to data mining and the use of geographic information systems.

      Honestly the category of digital humanities seems like it could do with being split into two (or several) smaller fields of study. I'm sure it already is, the same way ecology and ornithology are both biology, but at least with biology it can be summarized as "the study of life" - with digital humanities I still struggle to come up with something like that - "the study of anything that could possibly be explored further/easier with anything similar to a computer?"

    6. This alliance of geeks and poets has generated exhilaration and also anxiety. The humanities, after all, deal with elusive questions of aesthetics, existence and meaning, the words that bring tears or the melody that raises goose bumps. Are these elements that can be measured? Advertisement Continue reading the main story “The digital humanities do fantastic things,” said the eminent Princeton historian Anthony Grafton. “I’m a believer in quantification. But I don’t believe quantification can do everything. So much of humanistic scholarship is about interpretation.”

      Not to argue with the New York Times and a Princeton Historian, but are the digital humanities really limited to quantification? I think that's perhaps a bit of a narrow minded opinion, or that I'm misinterpreting. For example, digital art or the study of digital artwork could be considered digital humanities, and I don't think that digital art has anything to do with quantification.

      I do understand the idea that quantification and data can't "deal with elusive questions of aesthetics, existence and meaning, the words that bring tears or the melody that raises goose bumps" (frankly an awful sentence, but that's besides the point) without human interpretation. I've seen some truly horrifying or very encouraging statistics before that can evoke these responses like a piece of literature, but the statistics alone do not embody those reactions.

    1. I can see how the drama of this moment is enticing. It offers a grandeur, a sweeping purity to our possibly flawed and fumbling and ambivalent selves. It justifies all our failings and setbacks and mediocrities; it wasn’t us, it was men, or the patriarchy, holding us back, objectifying us. It is easier to think, for instance, that we were discriminated against than that our story wasn’t good enough or original enough to be published in The Paris Review, or even that it did not meet the editor’s highly idiosyncratic yet widely revered tastes. Or that a man said something awful and sexual to us while we were working on a television show, and we got depressed and could never again achieve what we might have. And yet do we really in our hearts believe that is the whole story? Is this a complete and satisfying explanation? There is, of course, sexism, which looms and shadows us in all kinds of complicated and unmappable ways, but is it the totalizing force, the central organizing narrative, of our lives? This is where the movement veers from important and exhilarating correction into implausibility and rationalization. (One of the deeply anonymous says, “This seems like such a boring way to look at your life.”)

      I absolutely agree with this conclusion--mob mentality has always been more detrimental than beneficial if at all, and we should be able to see this clearly in the United States today. However, I feel like this point could have been made in a satisfactory manner halfway through this essay. I could see this conclusion coming from the beginning of the second page and the bits about Lorin Stein and Moira Donegan's hypocrisy could have been a separate essay by themselves. Otherwise a very sensible and interesting read.

    2. I have a feeling that if one met @yoloethics or the rest of her Twitter cohort in person, they would seem normal, funny, smart, well read. But the vicious energy and ugliness is there beneath the fervor of our new reckoning, adeptly disguised as exhilarating social change. It feels as if the feminist moment is, at times, providing cover for vindictiveness and personal vendettas and office politics and garden-variety disappointment, that what we think of as purely positive social change is also, for some, blood sport.

      I love the references to the Roman empire here--the description of twitter followers as cohorts and the twitter frenzy as a blood sport really hits home the disturbing and vindictive nature of this entire issue.

    3. It can be hard to disentangle one man and the things he may or may not have done from hundreds of years of sexist oppression.

      In terms of workplace harassment in particular, this statement is revealing of a larger issue--I was searching for a comprehensive analysis of the global incidence of workplace sexual harassment in the, but apparently as of 2015 there was no survey done since 1994. This is truly alarming to me, as there is a sincere lack of data and this is just helping to feed this twitter frenzy issue. This may be the main reason why company policies are slow to change.

      Source: https://wol.iza.org/uploads/articles/188/pdfs/sexual-harassment-in-workplace.pdf

    4. “I like Lorin,” she told me. “I don’t have a personal stake in this.” She then informed me that he had sexually harassed two interns at Farrar, Straus and Giroux, where he had worked before his Paris Review tenure, leading to hushed-up, sealed settlements. She delivered this piece of highly specific information so confidently that I did not stop and think, even though I teach in a journalism department: Is this factually correct?

      I understand that this feeds into Roiphe's argument that the twitter frenzy happens the same way, but it would have served her argument better to include an instance from twitter itself. This and the next few paragraphs make this and the twitter issue seem resigned to human nature. The final paragraph is a good objection to it, but here its treated too lightly in my opinion.

    5. The night the New York Times broke the news of Stein’s resignation, I was with one of the deeply anonymous women in a coffee shop, and after I left she ran out and caught up to me on the dark street to tell me about it. When I got home, I saw that @MegaMoira had tweeted a photo of the piece with the words, “champagne anyone.” I thought of the email Lorin had sent me when my book on writers’ deaths, The Violet Hour, came out. It was such a strange, private project, but in a few lines he made it vivid again to me, renewed and energized me on a long winter afternoon to sit down and start something new. However one feels about the end of an era at The Paris Review, it doesn’t seem like a time for celebration.

      While I believe that Stein's resignation needed to happen, Roiphe is correct in her sentiment that nobody really wins from this situation. The Paris Review is worse off as a company for not having him anymore and the fact that he would commit such offenses broke the hearts of everyone close to him.

    6. However, after the list came out but before Lorin Stein resigned as editor of The Paris Review, she tweeted: “every profile of Lorin Stein calls him ‘skinny and bespectacled’ but here’s the thing: he’s not that skinny.” She added: “I guess ‘bespectacled, bald, and busting out [of] the bespoke shirts he’s still having made with 15 year old measurements’ doesn’t have the same ring to it.” Later, these tweets were deleted. But if we could think in less gendered terms for a moment, one could reasonably ask: Who is harassing whom?

      I absolutely agree here--this does not venture beyond petty name-calling and is more crucially a demeaning of the person based on his appearance, which is one of the behaviors considered harassment by Donegan. This certainly puts renewed insight into her mindset when she says she wants a more respectful world.

    7. I feel blessed to live in a society where you are free to walk through the city at night. I just don’t think those of us who are privileged white women with careers are really that afraid.

      The question of safety at night is an interesting one and I don't think the answer is as simple as the society (especially in the city, where intimacy is not ambient but compartmentalized) in which one acts. Even the presence of streetlights (and by extension the invention and proliferation of electric lighting) has a large impact on safety at night. Money surely has a place in it as well.

    8. Here is what the last few days have reminded me: white men, even those on the left, are so safe, so insulated from the policies of a reactionary presidency, that many of them view politics as entertainment, a distraction without consequences, in which they get to indulge their vanity by fantasizing that they are on the side of good. . . . The morning after the election, I found the penis-shaped shot glass in my kitchen and threw it against the wall. I am not proud of this, but it felt good to destroy something a white man loved.

      And yet, the way you write those sentences, especially the one at the end, makes it look like you are indeed proud of that. It makes you seem pettier than you intended to be. Also I really don't understand the concept of penis shaped shot glasses--it seems like it would be more difficult to drink from than a wide glass.

    9. Every woman, every day, when she leaves her house, starts to think about safety. Can I go here? Should I go out there?. . . Do I need to find a taxi? Is the taxi driver going to rape me? You know, women are so hemmed in by fear of men, it profoundly limits our lives.

      It really does not help Solnit's cause to make statements like this that are unverifiable in nature.

    10. In 1996, a six-year-old boy with Coke-bottle glasses, Johnathan Prevette, was suspended from school for sexual harassment after kissing a little girl on the cheek.

      Below is the source for the fact and the following passage. I agree with the sentiments of Johnathan's father in the report--I have seen many boys kiss girls on the cheek when I was in elementary school and this was blown completely out of proportion. It was obvious that the boy did not intend to cause that kind of harm, nor even conceive of it.

      https://www.nytimes.com/1996/09/27/us/6-year-old-s-sex-crime-innocent-peck-on-cheek.html

    11. Can this possibly be a good thing?

      This bit seems unnecessary to me. The entire previous part of this paragraph is devoted to the portrayal of whisper networks as consequences of bad things, so this would have served Roiphe's point better if it was at the beginning of the paragraph.

    12. For years, women confined their complaints about sexual harassment to whisper networks for fear of reprisal from men.

      I wanted to find a source for this information that is not a news story, but have failed so far.

    13. Before the piece was even finished, let alone published, people were calling me “pro-rape,” “human scum,” a “harridan,” a “monster out of Stephen King’s ‘IT,’?” a “ghoul,” a “bitch,” and a “garbage person”—all because of a rumor that I was planning to name the creator of the so-called Shitty Media Men list.
    14. No one would talk to me for this piece. Or rather, more than twenty women talked to me, sometimes for hours at a time, but only after I promised to leave out their names, and give them what I began to call deep anonymity.

      An example of toxic shame (or rather a fear of toxic shame as a result of the twitter culture as described by Roiphe in the following paragraphs) that is not necessarily present from childhood, but is certainly imposed by others.

    15. I am not trying to suggest that the list makers don’t understand the difference in scale between leering and assault, but rather that the blurring of common (if a little sleazy) behavior and serious sexual harassment reveals a lot about how they think. For them, the world is overrun with leering monsters you have to steer around, as if in a video game.

      It is precisely, in my opinion, because the writers knew the difference between assault and leering that they wrote the list in such a manner (specifically, a spreadsheet to be presented). It seems as though at least some of them were trying to hit somebody with it and that makes it a particularly disturbing example of refined cruelty to me.

    16. To do a close reading of the list: some of the offenses on the spreadsheet (“creepy DMs,” “weird lunch ‘dates,’” “leering,” “flirting,” “violent language,” and “leading on multiple women online”) seem not quite substantial or rare enough to put into the category of sexual misconduct. I am not even sure they merit a warning to a hopeful young employee. I have graduate students who go on to work for these sorts of publications, and I am very mother-hen-ish about them. But I can’t imagine sitting with one of my smart, ambitious students in my office, lined with shelves of books like The Second Sex and A Room of One’s Own and I Love Dick and The Argonauts, saying, “Before you go work there, I just want to warn you, that guy might leer at you.” I would worry I was being condescending, treating her like a child who doesn’t know how to handle herself in the world

      This seems very sensible to me. Anybody, I think, with a reasonable mindset would agree that adults are capable of dealing with such minor transgressions as flirting, leering, and violent language (whatever that means). Whether or not they treat such behavior as serious harassment is ultimately dependent on the context and on their particular judgment of the situation. This list seems to throw all meaningful context out the window and is very difficult to take seriously as a result.

    17. (“It feels Maoist,” says one of the deeply anonymous, while others question whether the list was ever designed to remain clandestine in the first place.)

      In my personal experience, spreadsheets are meant to be easily readable and organized in such a way that their purpose is to be a presentation of information and data to others. In other words, if this list was a spreadsheet, then I believe it was meant to be shared with as many people as possible and not really intended to be clandestine.

    18. The Shitty Media Men list, the anonymously crowd-sourced spreadsheet chronicling sexual misconduct in the publishing world, is a good example. If we think of how we would feel about a secretly circulating, anonymously crowd-sourced list of Muslims who might blow up planes, the strangeness of the document snaps into focus.

      A very apt comparison. Writers of both documents would be susceptible to prosecution in the form of libel suits regardless of the degree of truth behind the allegations. In fact, one libel suit was filed by Stephen Elliott against Moira Donegan. Elliott also wrote a response to the Shitty Media Men List;

      Suit: https://www.latimes.com/books/la-et-jc-stephen-elliott-20181015-story.html

      Response: https://quillette.com/2018/09/25/how-an-anonymous-accusation-derailed-my-life/

    19. (One of the editors of n+1, Dayna Tortorici, tweets: “I get the queasiness of no due process. But . . . losing your job isn’t death or prison.”)

      Ironically, this impatience with due process would violate basic human rights as codified in the 1949 Geneva conventions and would doubly violate the United States bill of rights.

      https://ihl-databases.icrc.org/customary-ihl/eng/docs/v2_rul_rule100

    1. Sonicated cells of E. coli having recombinant vector was centrifuged. Supernatant was dispensed into 0.2 % v/v xylan agar plate and incubated for 4 h. The plates were then flooded with Congo red solution (0.2 % w/v) for 30 min and destained with 1M NaCl solution till a clear zone of xylan hydrolysis was visible. The plates were gently shaken on a shaker to accelerate the process of staining/destaining
    2. Qualitative detection of xylanolytic activity by plate assay
    3. Preparation of electrocompetent cells (E. coli cells) A protocol was employed. The procedure was carried out in cold under sterile conditions as follows: •A single colony of E. coli DH10B/ DH5α/XL1blue was inoculated in 20 mL of LB medium and grown overnight at 30 °C. •500 mL LB medium was inoculated with 5mL of this overnight grown culture of the E. coli and incubated with vigorous shaking (250 rpm) at 30 °C until an A600of 0.5 - 0.8 was achieved. •The cells were chilled in ice for 10-15 min and transferred to prechilled Sorvall® centrifuge tubes and sedimented at 4,000 rpm for 20 min at 4 °C. •The supernatant was decanted and cells were resuspended in 500 mL of sterile ice-cold water, mixed well and centrifuged as described above. •The washing of the cells described above was repeated with 250 mL of sterile ice-cold water, following which cells were washed with 40 mL of ice-cold 10 % (v/v) glycerol and centrifuged at 4,000 rpm for 10 min. •The glycerol solution was decanted and the cell volume was recorded. The cells were resuspended in an equal volume of ice-cold 10 % glycerol. •Cells were then dispensed in 40 μL volumes and stored at -80 °C until required.
    4. Electrotransformation
    5. The metagenomic DNA extracted from above defined protocol was digested with Sau3A1 at conditions optimized to generate maximum fragment in the size range of 2-6 kb. Different concentration (0.05 to 1 unit) of enzyme was used to optimize the digestion of 1 μg of DNA. Reactions were carried out in a final volume of 30 μl each in an Eppendorf of 1.5 mL. Reaction mixture (1 μg DNA having 3 μL NEB buffer 3 and 0.3 μL of 10X BSA) were kept at 37 °C for 10 min and stopped by heat inactivation at 80 °C for 20 min. Different digested reactions were checked for the desired fragments using 0.8 % (w/v) agarose gel electrophoresis. After optimization of DNA fragments for the appropriate size, a large scale digestion was carried out and the fragments (2-8 kb) were purified from low melting agarose gel using gel extraction method according to the manufacturer’s protocol (Qiagen gel extraction kit, Germany)
    6. Insert DNA preparation
    7. Purity of the DNA extracted from various environmental samples was confirmed by subjecting the extracted DNA to restriction digestion. DNA was digested with Sau3AI (New England Biolabs). One μg of metagenomic DNA in 20 μL reaction mixture was treated with 0.5 U of Sau3AI and incubated at 37 °Cfor 10 min. The reaction was terminated at 80 °C for 20 min and the digested DNA was fractionated on 1.2 % (w/v) agarose gel.
    8. Restriction digestion
    9. The isolated DNA was diluted (1:100) with MQ. The concentration (mg mL-1) of the DNA [N] was determined spectrophotometrically by recording absorbance at 260 nm (A260) as: A260 = ε 260[N]where ε 260 is the extinction coefficient of DNA (50 for ds DNA) [N] = concentration (mg mL-1) of DNA The concentration of ds DNA [N] was calculated as [DNA] (mg mL-1) = A260/ε 260 [DNA] (μg mL-1) = A260 × 50 × dilution factor Purity of DNA was checked by measuring absorbance at 260 and 280 nm and calculating the A260/A280 ratio (Sambrook et al., 1989). A DNA sample was considered pure when A260/A280 ranged between 1.8-1.9. An A260/A280 < 1.7 indicated contamination of the DNA preparation with protein or aromatic substances such as phenol, while an A260/A230 < 2.0 indicated possible contamination of high molecular weight polyphenolic compounds like humic substances.
    10. Determination of DNA quantity and purity
    11. as well as commercial methods (MN kit, Germany; Mo-Bio kit, CA, USA; Zymo soil DNA kit, CA, USA) according to the manufacturer’s protocols and compared in terms of DNA yield and purity.
    12. The soil DNA from Pantnagar and Lonar soil samples were also extracted by various manual (Desai and Madamwar, 2007; Agarwal et al., 2001; Yamamoto et al., 1998
    13. Alternatively metagenomic DNA was extracted from the alkaline soil samples by using different commercial kits (UltraClean™, PowerSoil™ [Mo Bio Laboratories Inc., Carlsbad, CA, USA], Nucleospin kit [Macherey-Nagal, Germany] and Zymo soil DNA isolation kit [CA, USA]). The DNA was finally suspended in 100 μL of sterile Milli Q water for further analysis.
    14. Commercial kits
    15. Comparison of yield and purity of crude DNA
    16. Soil (1 gm) was suspended with 0.4 gm (w/w) polyactivated charcoal (Datta and Madamwar, 2006) and 20 μL proteinase K (10 mg mL-1) in 2 mL of modified extraction buffer [N,N,N,N cetyltrimethylammonium bromide (CTAB) 1% w/v, polyvinylpolypyrrolidone (PVPP) 2% w/v, 1.5 M NaCl, 100mM EDTA, 0.1 M TE buffer (pH 8.0), 0.1M sodium phosphate buffer (pH 8.0) and 100 μL RNaseA] [Zhou et al., 1996] in 20 mL centrifuge tubes to homogenize the sample and incubated at 37 °C for 15 min in an incubator shaker at 200 rpm. Subsequently, 200 μL of 10% SDS was added to the homogenate and kept at 60 °C for 2 h with intermittent shaking. DNA was precipitated by adding 0.5 V PEG 8000 (30 % in 1.6 M NaCl) and left at room temperature for an hour (Yeates et al., 1998). The precipitated DNA was collected by centrifugation at 8000 x g at 4 °C. The supernatant was discarded and pellet was dissolved in 1 mL of TE buffer (pH 8.0) and then100 μL of 5 M potassium acetate (pH 4.5) was added and incubated at 4 °C for 15 min. The supernatant was collected after centrifugation at 8000 x g and treated with equal volumes of phenol: chloroform (1:1) followed by chloroform: isoamylalcohol (24:1) at 8000 x g for 15 min
    17. PROTOCOL FOR OPTIMIZATION OF HUMIC ACID-FREE DNA FROM ALKALINE SOILS
    18. Various strains of Escherchia coli (DH5α, XL1Blue, DH10B) were used as hosts for the propagation of recombinant vectors. In addition, Bacillus subtilis was used as a host for the expression of xylanase gene from the recombinant vector pWHMxyl. Different vectors used in this investigation are listed in
    19. BACTERIAL STRAINS
    20. Soil, sediment, effluent, and water samples have been collected from various hot and alkaline regions of India and Japan in sterile polyethylene bags/bottles. The samples were transported to the laboratory and preserved at 4 °C. Temperature and pH of the samples was recorded.
    21. COLLECTION OF SAMPLES
    1. or DNA isolationJrom P. Jalciparum, genomic DNA kit from Qiagen (Germany) was used. Isolation was done following manufacturer's instructions. Briefly, infected erythrocytes (5 ml at 10% parasitemia) were centrifuged at 3,000 g for 2 min. The cells were washed once in cold PBS and resuspended in 1 ml. Following which, 10 ilL of 5% saponin (final concentration 0.05%) was added and' mixed gently. After lysis, the mix was immediately centrifuged at 6,000 g for' 5min. Further steps were, carried out according to the manufacturer's instructions to isolate genomic DNA. DNA was quantified by measuring absorbance at 260 nm I using a UV -spectrophotometer
    2. Genomic DNA Isolation from Parasite Culture
    3. lasmodium Jalciparum strain 3D7 (MR4, American Type Culture Collection) was i used for all the experiments except where gametocyte rich culture was required. I For generating gametocytes, 3D7 A a variant of 3D7 was used. The parasite was cultured as describerl below:
    4. lasmodiumfalciparum culture