32 Matching Annotations
  1. Last 7 days
    1. I went with them out to the veranda. On the green Sound, stagnant in the heat, one small sail crawled slowly toward the fresher sea. Gatsby’s eyes followed it momentarily; he raised his hand and pointed across the bay.

      Daisy and Gatsby liked each other and they cared for each other.

    2. Gatsby, who represented everything for which I have an unaffected scorn. If personality is an unbroken series of successful gestures, then there was something gorgeous about him, some heightened sensitivity to the promises of life, as if he were related to one of those intricate machines that register earthquakes ten thousand miles away.

      She thinks that Gatsby is a bit charming.

    3. I began to have a feeling of defiance, of scornful solidarity between Gatsby and me against them all.

      This sentence shows how fully Nick aligns himself with Gatsby in death, even more than in life. He feels a moral loyalty that the rest of Gatsby’s world refuses to offer. It also marks Nick’s final separation from the careless society he has been observing.

    4. “I feel far away from her,

      This sentence shows Gatsby’s growing fear that his dream cannot survive real contact with Daisy. He realizes that she cannot see the world he has built around her. It also exposes the emotional distance that his fantasy tried to hide.

    5. he revalued everything in his house according to the measure of response it drew from her well-loved eyes.

      This sentence shows how deeply Gatsby bases his self-worth on Daisy’s approval. His possessions stop being symbols of success and become tools for winning her reaction. It also reveals how unstable his dream is, because it depends entirely on her gaze.

    1. Specific words and images make your writing clearer, more precise, and often more interesting. Whenever possible, avoid overly general words in your writing; instead, try to replace general language with particular nouns, verbs, and modifiers that convey details and that bring yours words to life. Add words that provide color, texture, sound, and even smell to your writing.

      This makes me realize that using specific words could make such a big difference.Adding small details like color or sound can suddenly make writing feel more alive.

  2. Mar 2025
  3. southtexascollege.blackboard.com southtexascollege.blackboard.com
    1. This flea is you and I, and this Our marriage bed, and marriage temple is;

      The speaker tells the listener that the flea signifies their marriage bed and temple. By this logic, it sets the two being already married. By making the flea play the part of a temple and bed, it seemingly makes this small, insignificant bug, play a huge role within the poem.

  4. May 2024
  5. Jan 2022
  6. Mar 2021
  7. Jul 2019
  8. Jun 2019
  9. May 2019
    1. eppendorf tube was put at the bottom of the column to collect the eluate. The column was respun as before and the purified probe collected in the eppendorf tube, the unincorporated nucleotides remaining within the column. One ul aliquot from the purified probe was diluted 100 fold, mixed well and 1 ul aliquots were put in triplicate into 3 ml scintillation fluid containing vials which were counted in a Beckman Liquid Scintillation Counter. The total radioactivity of the probe was calculated by multiplying the mean radioactivity of the three diluted samples with a factor of 104 ( dilution factor, 102, total reaction volume, 102 ). The specific activity of the probes ranged from 1 X 107 to 5 x·1o7 cpm 1 ug DNA. The probe purified by the above method did not require any further purification.
    2. The nick translated probe was purified by a spun column procedure to remove the unincorporated nucleotides. A sterile 1 ml syringe was plugged at the lower end with siliconised glass wool. The syringe was then filled with Bio-gel P-4 Bio Rad Laboratories, USA ) equilibrated in advance with TE. For doing this, 30 grammes of Bio-gel P-4 was slowly added into 250 ml of TE ensuring a good dispersion of the powder. This was then autoclaved at 15 psi for 20 minutes. After cooling, the supernate was decanted and replaced with an equal volume of sterile TE. The slurry was stored at 4°C. The slurry was poured upto the 1 ml mark in the syringe. The syringe was placed into a centrifuge tube and spun at 2000 · rpm for 3 minutes. The column was packed by repeating this process till the packed column volume reached 1 ml mark. Next, 50 ul of 2 mg 1 ml denatured salmon sperm DNA was loaded as carrier and the column spun as before. 100 ul of TE was then added to the column and it was respun as before. Finally, the nick translation reaction was diluted to 100 ul with TE and loaded on to the column. A sterile 1.5 ml
    1. 10 min at 4 ̊C and gently resuspended in 20 ml ice-cold Inoue transformation buffer. To this cell suspension, 1.5 ml sterile DMSO was added and swirled gently. Cell suspension was kept on ice for 10 min and 50 μl volume was aliquoted to chilled sterile microcentrifuge tubes. Cells were immediately snap-frozen in liquid nitrogen and stored at -80 ̊C
    2. A single colony of E. coli DH5α strain was inoculated in 10 ml LB medium and incubated at 37 ̊C for overnight. 4 ml overnight culture was inoculated in 2 lt SOB medium and incubated at 18 ̊C till the OD600 reached to 0.5. Cultures were centrifugedat 2,500 g for 10 min at 4 ̊C and harvested cells were washed gently with80 ml ice-cold Inoue transformation buffer. Cells were collected by centrifugation at 2,500 g for
    1. respectively. The resulting constructs pRR14 and pRR15 were transferred in E.coliDH5α. Through triparental mating using pRK600 helper plasmid the construct were transferred in E.coliS17-1. After confirming pRR14 and pRR15 constructs by sequencing, the constructs were then introduced into BXOR1 strain through biparental mating using E. coliS17-1. X. oryzaepv. oryzicolaGUS and GFP reporter strains were selected on PS medium plates containing suitable antibiotics. Since pVO155 cannot replicate in X. oryzaepv. oryzae, ampicillin and kanamycin-resistant colonies were obtained upon chromosomal integration of the plasmid using the cloned DNA sequence as a region of homology. pProbeGTcan replicate independently in Xanthomonasand report for the gene expression
    2. Glucuronidase (GUS) reporter gene fusion and GFP reporter fusion was created by using the suicide plasmid pVO155 having a promoterless gusAgene (Oke and Long, 1999), and pProbeGThaving a promoterless GFP (Miller et al., 2000). To construct the xsuA::gusAand xsuA::gfptranscriptional fusion, a 611-bp DNA fragment containing the putative promoter of the xssoperon (+213 to −398) was amplified by using the SCRsid_ pProbeGFP_F and SCRsid_ pProbeGFP_R primers (Table 2.2). This promoter fragment was subsequently digested with HindIII and BamHI,and directionally cloned upstream of the promoterless gusAand gfpgene in pVO155 and pProbeGTplasmids to create the xsuA::gusAand xsuA::pProbeGT(gfp) reporter constructs pRR1
    1. lines were used for stable knockdown of IP6K1 expression. Viral particles harboring either non-targeting control or IP6K1directed shRNA were used to infect HeLa or HCT116 cell lines at 0.5 MOI, following treatment with polybrene (8 μg/mL)for 2 h.After 48 h, transduced cells were selected with 2 μg/mL puromycin. Medium was changed twice a week and observed for colony formation. After reaching the optimum growth, selected cells were maintained in DMEM supplemented with 10% FBS and 1 μg/mL puromycin as