desired temperaturefor 45 minutes and plated on an appropriate selective medium at various dilutions. An aliquot of cell suspension to which plasmid DNA was not added served as a negative control. B. Inoue method i. Preparation of high efficiency competent cells Competent cells for high efficiency transformation were prepared by the method of (Inoueet al., 1990)with few modifications. An overnight culture of the strain (routinely DH5α) was subculturedinto fresh sterile LB broth in 1:100 dilutions and grown at 18ºC to anA600of 0.55. The cells were harvested by centrifugation at 2500rpm for 10 minutes at4ºC. Thesecells wereresuspended in0.4 volumes of INOUE buffer andincubated in ice for 10 minutes. The cells were recovered by centrifugation at 2500rpm at 4ºC for 10 minutes and finally resuspended in 0.01 volume of the same buffer. Sterile DMSO was added to a final concentration of 7%. After incubating for 10 minutes in ice, the cells were aliquoted in 100μl volumes, snap frozen in liquid nitrogen and stored at –80ºC. ii. Transformation protocolFor transformation, the required number of vials wasthawed on ice and the transformation protocol as described for CaCl2method was employed

A. Calcium chloride(CaCl2)method For routine plasmid transformation, following method which is a modification of that described by(Cohenet al.,1972)was used. An overnight culture of recipient strain was subcultured 1:100 in fresh LB medium and grown till mid-exponential phase. The culture was chilled on ice for 20 minutes, and the steps thereafter performed at 4ºC. 10 ml of culture was centrifuged and pelletwas resuspended in 5 ml of 0.1M CaCl2. After 5 minutes of incubation on ice, the cells were again centrifuged and resuspended in 1ml of 0.1M CaCl2. The suspension was incubated onice for 45 minutes. To the 100μl aliquot of the cellsuspension plasmid DNA (20-200ng in less than 10μl volumes) was added, incubated for 30-40 minutes on ice and given a heat shock for 90 seconds at 42ºC. The cultures were rapidly chilled for 1 minute, mixed with 0.9ml of LB broth and incubated at

Transformation

To 2 ml of fresh overnight culture of recipient strain, 108 pfu equivalent of phage lysate was added and incubated at 37ºC without shaking for 30 minutes to facilitate phage adsorption. The unadsorbed phage particles were removed by centrifugation at 6000 rpm for 5 minutes and the pellet ofbacterial cells was resuspended in 5 ml of LB broth containing 20 mM sodium citrate to prevent further phage adsorption. This was incubated for 25-60 minutes at desired temperaturewithout shaking to allow the phenotypic expression of the antibiotic resistance gene. The mixture was then centrifuged and the pellet was resuspended in 300 μl of 0.1M citrate buffer. 100 μl aliquots were spreadon appropriate antibiotic containing plates supplemented with 2.5 mM sodium citrate. A control tube without addition of P1 lysate was also processed in the same way. In the case of selection of nutritional requirement, the infection mixture was centrifuged, resuspended in 300 μl of 0.1M citrate buffer and plated without phenotypic expression

10–6)were mixed with 0.1 ml of fresh culture grown in Z-broth. After 30 min of adsorption at 37ºC without shaking, each mixture was added on a soft agar overlay of Z-agar plates and incubated overnight at 37ºC. The phage titrewas calculated from the number of plaques obtained on the plates as follows: Phage titre(pfu) per ml = No. of plaques ×dilution factor ×1000/vol.of lysate added (in μl)

Phage P1 lysate preparation by broth method

Phage P1 transduction

0.3 ml of overnight culture of the donor strain in Z-broth was mixed with 107plaque forming units (pfu) of a stock P1 phage lysate prepared on strain MG1655. Adsorption was allowed to occur at 37ºC for 30 minutes and the lysate was prepared by broth method. To 0.3 ml of infection mixture, 8-10 ml of Z-broth was added and incubated at 37ºC with slow shakinguntil the visible lysis of the culture occurred (in 4-6 hours). The lysate was treated with 0.2ml of chloroform, centrifuged and the clear lysate wasstored at 4ºC with chloroform.Quantitation of Plaque forming units (pfu)To quantitate the titreof P1 lysate preparation, titration was done using P1 phage sensitive indicator strain such as MG1655. 100 μleach of dilution of phage (typically 10–5,

Genetic Techniques

We performed some manipulation checks to examine the internal validity of the perceptual-cognitive skill tests and any learning effects as a result of watching the same video clips multiple times

Four groups of subjects were compared in the study.

(1)studies relating clinical focal frontal lobe disor-ders to violent behaviour; (2) studies reportingneuropsychological measures of frontal lobefunction in aggressive and antisocial subjects;(3) studies of clinical neurological findings inviolent and criminal populations; and (4) neu-roimaging studies of aggressive and violentsubjects

Methods

Second, we recruited a group of college students majoring in chemistry tocompletetheprocedure

Weemployed two control groups as a performance standard. First, as in our pre-vious study, we asked a group of people to fill out the questionnaire withoutproviding them any information whatsoever about the specific case except itstype (i.e., murder).

First, a group of senior police detectives who possessedsubstantial experience in criminal investigations participated. Second, werecruited a group of detectives who specialized in the investigation of homi-cide. Third, to contrast investigative experience to general experience as apolice officer, a sample of trainee detectives who possessed significant expe-rience in police general duties but only a marginal amount of experience incriminal investigations was obtained. Finally, a sample of police recruits wasalso used.

Our research took place at Florida State University (FSU), a large, flag-ship, Research–1 institution in the Southeast US, had IRB approval and spanned two semesters totaling thirty-five weeks, over the fall of 2009 and the spring of 2010.

Motif search, calculation of motif activities, and target prediction

I buy into Newton’s philosophy that we see further by standing on the shoulders of giants.

Instead of being simply the recipient of negative comments, the reader, in this situation, sees an alternative possibility.  It's not the only one.  It's another option on the way to success. 

Improved Protocols for Illumina Sequencing

machine learning

A practice is included in our list if large numbers of researchers use it and large numbers of people are still using it months after first trying it out. We include the second criterion because there is no point in recommending something that people won't actually adopt.

applying networkand content analyses

I have written elsewhere: ‘The way of becoming actually free is both simple and practical. One starts by dismantling the sense of identity that has been overlaid, from birth onward, over the innate self until one is virtually free from all the social mores and psittacisms. Virtually free from all the beliefs, ideas, values, theories, truths, customs, traditions, ideals, superstitions ... and all the other schemes and dreams. One can become aware of all the socialisation, of all the conditioning, of all the programming, of all the methods and techniques that were used to produce what one thinks and feels oneself to be – a wayward identity careering around in confusion and illusion. A ‘mature adult’ is actually a lost, lonely, frightened and very cunning entity. However, it is never too late to start in on uncovering and discovering what one actually is. One can become virtually free from all the insidious feelings – the emotions and passions – that fuel the mind and give credence to all the illusions and delusions and fantasies and hallucinations that masquerade as visions of The Truth. One can become virtually free of all that which has encumbered humans with misery and despair and live in a state of virtual freedom ... which is beyond normal human expectations anyway. Then, and only then, can the day of destiny dawn wherein one becomes actually free. One will have obtained release from one’s fate and achieved one’s birthright ... and the world will be all the better for it’.

This article can be utilized as an argument piece wherein I can help build on my argument.

mechanistic approach

results of your research, and within the context of the study, we found ample evidence about significant cha

essay has a goal of bringing

The point of replication isn't to shame researchers — it's to build better science

Performance CategoryDesign Categoriesi. StructureFrame design, shape and materials –for functionii. MobilityThrusters: number, power, orientationiii. SensorsCameras, lights, sonar, touch sensors, compass, GPSiv. ToolsArms, claws, rakes, wrenches, hammersv. Ranging DistanceTether length: waterproofing required vi. Buoyancy/ BallastFixed or variable, location and materialsvii. ControlsRC via wire or signal via fibre optic cableviii. Other?Depends on the specific mission

For the humanities syllabi, I also asked how many tools were being taught in each course.

I looked at whether the courses: 1) required a collaborative project and 2) set aside time to discuss the challenges of collaboration or cross-disciplinary research (or had readings that indicated such).

With each syllabus I looked for general focus (is it tools-, training-, or topics-focused?), the breadth of assigned readings (is the literature from librarianship, humanities fields, or both?), and the structure of project(s) (collaborative, individual).

formative study

craft cultures

The models criticized earlier do not need to be trashed. They are not just plain wrong. It's just that their sphere of applicability must be recognized as limited to a particular mode of exis­ tence, or a particular dimension of the real (the degree to which things coincide with their own arrest).

iterative-inductive research (that evolves in design through the study), drawing on a family of methods,

'rbe concepts appear and reappellT like II revolving cast of characters, joining forces or interfer ing with each other in a tumble of abstract intrigues-at rimes (I admit) barely controlled

x = -a +/- (a2 -b)1/2

get information of the literature’s abstr

vironment’,’trade’,’trad

In use these should be united, so that the same oration can expound purely, speak ornately, and express thought wisely. However, the precepts of pure diction, ornate delivery, and intelligent treatment must be kept separate and should not be confused.

method

Moreover, we must not even trust to the first learning by heart; it will be better to have syllables repeated and to impress them long upon the memory; and in reading too, not to hurry on, in order to make it continuous or quick, until the clear and certain connection of the letters become familiar, without at least any necessity to stop for recollection

false knowledge

SOCRATES: And that, Gorgias, was what I was suspecting to be your notion; yet I would not have you wonder if by-and-by I am found repeating a seemingly plain question; for I ask not in order to confute you, but as I was saying that the argument may proceed consecutively, and that we may not get the habit of anticipating and suspecting the meaning of one another's words; I would have you develope your own views in your own way, whatever may be your hypothesis. GORGIAS: I think that you are quite right, Socrates.

Now I want to know about rhetoric in the same way;—is rhetoric the only art which brings persuasion, or do other arts have the same effect? I mean to say—Does he who teaches anything persuade men of that which he teaches or not?

And here let me assure you that I have your interest in view as well as my own