User contributions

From 2012.igem.org

For Ajl58 (talk | block log | logs)

(Latest | Earliest) View (newer 50 | older 50) (20 | 50 | 100 | 250 | 500)

23:40, 3 October 2012 (diff | hist) N User:DrJones1935/1 October 2012 ‎ (Created page with "====I. Re-check plates that were allowed to grow more overnight==== =====RESULTS:===== No appreciable change in E. coli plates. 2 blue colonies developed on the ADP1 1 plate and...") (top)
23:40, 3 October 2012 (diff | hist) N User:DrJones1935/30 September 2012 ‎ (Created page with "====I. Check Transformation Plates for Colonies==== =====RESULTS:===== *''B. subtilis'' PY79 - no colonies on any plates *''E. coli'' MachI {| class="wikitable" |- ! Plate !! ...")
23:39, 3 October 2012 (diff | hist) N User:DrJones1935/29 September 2012 ‎ (Created page with "====I. Inoculate Liquid Cultures for Transformation==== *For MachI, ADP1, and PY79 cultures, inoculate 10 mL of LB with 100 uL of overnight cultures *For EcNR1 and EcNR2ΔlacZ, ...") (top)
23:39, 3 October 2012 (diff | hist) N User:DrJones1935/10 September 2012 ‎ (Created page with "====I. Measure the Concentration of Miniprepped Plasmids==== *Bring samples and Buffer EB bottle upstairs to Take3 plate reader *Add 2 uL of each sample according to the followi...")
23:39, 3 October 2012 (diff | hist) N User:DrJones1935/6 September 2012 ‎ (Created page with "====I. Check Overnight Cultures for Growth==== *RESULTS: All cultures showed growth. ====II. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.4 g of agarose and add to...") (top)
23:38, 3 October 2012 (diff | hist) N User:DrJones1935/5 September 2012 ‎ (Created page with "====I. Check Old Plates for Growth==== *RESULTS: There was XL1-Blue colony growth on Kan50 plates from 15 Aug 2012 that was not present on controls for the 1:1 (~4 colonies) and...") (top)
23:38, 3 October 2012 (diff | hist) N User:DrJones1935/15 August 2012 ‎ (Created page with "====I. Measure the Concentration of Extracted DNA==== *Bring samples and water upstairs to Take3 plate reader *Add 2 uL of each sample according to the following chart: {| class...")
23:37, 3 October 2012 (diff | hist) N User:DrJones1935/14 August 2012 ‎ (Created page with "====I. Check Plates for Growth==== ''RESULTS:'' Growth on positive controls was a large number of colonies but not a lawn as expected. No growth on any of the <span style="colo...") (top)
23:37, 3 October 2012 (diff | hist) N User:DrJones1935/13 August 2012 ‎ (Created page with "====I. Have Jason Repeat Experiment from Saturday with 10, 20, and 30 ug/mL Tetracycline Plates==== ====II. Design Primers for pBAV1K to Check Orientation of Beta Homolog Librar...") (top)
23:36, 3 October 2012 (diff | hist) N User:DrJones1935/11 August 2012 ‎ (Created page with "====I. Ligate Digested Plasmid and Digested Cassette Together==== *1:1 vector:insert ratio - Mix: :*0.34 uL water :*2 uL 10X T4 Buffer :*6 uL plasmid DNA (~0.05 pmol) :*10.66 uL...") (top)
23:36, 3 October 2012 (diff | hist) N User:DrJones1935/10 August 2012 ‎ (Created page with "====I. Digest pBAV1K-T5-gfp Plasmid and Cassette with XbaI and PstI==== *Mix (plasmid): :*37.5 uL water :*5 uL pBAV1K-T5-gfp DNA (~1 ug at 200 ng/uL) :*0.5 uL 100x BSA :*5 uL 10...")
23:35, 3 October 2012 (diff | hist) N User:DrJones1935/9 August 2012 ‎ (Created page with "====I. Prepare for Gel Extraction of Cassette==== *'''Make Gel:''' :*Measure out 0.40668 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix...")
23:35, 3 October 2012 (diff | hist) N User:DrJones1935/8 August 2012 ‎ (Created page with "====I. Prepare for Gel Extraction of LacZ Product from HF2==== *'''Make Gel:''' :*Measure out 0.40175 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and...")
23:34, 3 October 2012 (diff | hist) N User:DrJones1935/7 August 2012 ‎ (Created page with "====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.50131 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix :*Cover flask with ...") (top)
23:33, 3 October 2012 (diff | hist) N User:DrJones1935/6 August 2012 ‎ (Created page with "====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.50131 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix :*Cover flask with ...") (top)
23:33, 3 October 2012 (diff | hist) N User:DrJones1935/5 August 2012 ‎ (Created page with "====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.50325 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix :*Cover flask with ...") (top)
23:32, 3 October 2012 (diff | hist) N User:DrJones1935/4 August 2012 ‎ (Created page with "====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.45035 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix '''(0.9% agarose ge...") (top)
23:32, 3 October 2012 (diff | hist) N User:DrJones1935/3 August 2012 ‎ (Created page with "====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.40136 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix '''(0.8% agarose ge...") (top)
23:30, 3 October 2012 (diff | hist) N User:DrJones1935/2 August 2012 ‎ (Created page with "====I. Prepare for Gel Extraction of a PCR Product from 27 Jul 2012==== *'''Make Gel:''' :*Measure out 1.00721 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE ...")
23:29, 3 October 2012 (diff | hist) N User:DrJones1935/1 August 2012 ‎ (Created page with "====I. Order Beta Homolog Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm (Tm overhang/Tm anneal)...") (top)
23:29, 3 October 2012 (diff | hist) N User:DrJones1935/30 July 2012 ‎ (Created page with "====I. Prepare for Gel Extraction of a PCR Product from 27 Jul 2012==== *'''Make Gel:''' :*Measure out 0.75 g of agarose and add to a 250 mL E-flask :*Add 75 mL of 0.5x TBE buf...")
23:28, 3 October 2012 (diff | hist) N User:DrJones1935/29 July 2012 ‎ (Created page with " ====I. AGE of PCR Samples==== *'''Make Gel:''' :*Measure out 0.5 g of agarose and add to a 250 mL E-flask :*Add 50 mL of 0.5x TBE buffer and swirl to mix :*Cover flask with a ...") (top)
23:28, 3 October 2012 (diff | hist) N User:DrJones1935/27 July 2012 ‎ (Created page with "====I. Cross-over PCR==== *'''Template Mix Specifications:''' {| class="wikitable" |- ! Template !! Size (bp) !! Amount Wanted (pmol) !! Amount Wanted (ng) !! Concentration (ng/...") (top)
23:27, 3 October 2012 (diff | hist) N User:DrJones1935/26 July 2012 ‎ (Created page with "====I. Send Samples for Sequencing==== *Mix samples according to Keck Guidelines {| class="wikitable" |- ! Sample Name !! Water!! DNA !! F primer !! R primer !! Barcode # |- | L...") (top)
23:27, 3 October 2012 (diff | hist) N User:DrJones1935/24 July 2012 ‎ (Created page with "====I. Order Sequencing Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm !! Length (bp) !! %GC ...") (top)
23:27, 3 October 2012 (diff | hist) N User:DrJones1935/23 July 2012 ‎ (Created page with "====I. Run Diagnostic PCR on Sample 3==== *'''Mix Reagents:''' :*Mix according to the following table {| class="wikitable" |- ! !! A !! B !! C !! D !! E !! F |- ! Reagent !!...")
23:25, 3 October 2012 (diff | hist) N User:DrJones1935/21 July 2012 ‎ (Created page with "====I. PCR==== *'''Mix Reagents:''' :*Mix according to the following table, where 2 = LacZ, 3 = CAT*, 4 = tetr, 0.1-0.3 = +template, and 0.4 = -template ::*For exampl...") (top)
23:25, 3 October 2012 (diff | hist) User:DrJones1935/22 July 2012 ‎ (→I. PCR) (top)
23:23, 3 October 2012 (diff | hist) N User:DrJones1935/22 July 2012 ‎ (Created page with "====I. PCR==== *'''Mix Reagents:''' :*Mix according to the following table, where 2 = LacZ, 3 = CAT*, 4 = tetr, 0.1-0.3 = +template, and 0.4 = -template ::*For exampl...")
23:23, 3 October 2012 (diff | hist) N User:DrJones1935/19 July 2012 ‎ (Created page with "====I. Measure the Concentration of Miniprepped Plasmids==== *Bring samples and Buffer EB bottle upstairs to Take3 plate reader *Add 2 uL of each sample according to the followi...")
23:22, 3 October 2012 (diff | hist) N User:DrJones1935/18 July 2012 ‎ (Created page with "====I. Agarose Gel Electrophoresis (AGE) of PCR Samples==== *'''Make Gel:''' :*Measure out <strike>0.5 g</strike> 0.50497 g of agarose and add to a 250 mL E-flask :*Add 50 mL o...") (top)
23:22, 3 October 2012 (diff | hist) N User:DrJones1935/17 July 2012 ‎ (Created page with "====I. Dilute primers to 10 uM working stock==== *Mix: :*2 uL 200 uM primer stock :*38 uL sterile, DNAse-, RNAse-free dH2O *''Note: I think that the p5 primers got mi...") (top)
23:21, 3 October 2012 (diff | hist) N User:DrJones1935/16 July 2012 ‎ (Created page with "====I. Ordered Cross-over PCR Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm (Tm overhang/Tm ann...")
23:21, 3 October 2012 (diff | hist) N User:DrJones1935/15 July 2012 ‎ (Created page with "====I. Check Cultures for Growth==== ''RESULTS (~09:30)'': *Overnight cultures of plasmid strain pBAV1K-T5-gfp both grew robustly overnight. I will not centrifuge them together ...") (top)
23:19, 3 October 2012 (diff | hist) N User:DrJones1935/13 July 2012 ‎ (Created page with "====I. Check Plates for Growth==== *''RESULTS'': All overnight plates of ''B. subtilis'' showed growth, including the plates from two days ago left on the bench top. I have saved...") (top)
23:18, 3 October 2012 (diff | hist) User:DrJones1935/12 July 2012 ‎ (→I: Check WT B. subtilis Liquid Culture for Growth) (top)
23:17, 3 October 2012 (diff | hist) N User:DrJones1935/11 July 2012 ‎ (Created page with "====I. Check Plates and Cultures for Growth==== ''RESULTS (~09:45)'': *All plates showed growth except ''A. baylyi'' ADP1''ΔmutS''. ECNR2 plate was sparse, but had colonies. Al...") (top)
23:16, 3 October 2012 (diff | hist) N User:DrJones1935/10 July 2012 ‎ (Created page with "====I. Streak cells for upcoming transformation with pBAV1K-T5-gfp plasmid==== *Using sterilized inoculating loop, scrape top layer of ice of glycerol stock and streak cells ont...") (top)
23:15, 3 October 2012 (diff | hist) N User:DrJones1935/5 July 2012 ‎ (Created page with "====I. Ordered Cross-over PCR Primers from Keck==== *Primers were ordered with the name/sequence in the following table: {| class="wikitable" |- ! Name !! Tm (Tm overhang/Tm an...") (top)
23:14, 3 October 2012 (diff | hist) N User:DrJones1935/30 June 2012 ‎ (Created page with "====I. Check overnight cultures for growth==== *As of ~2:30 PM when I got in, both cultures were cloudy corresponding to saturation ====II. Make 60% glycerol stock from 100% st...") (top)
23:14, 3 October 2012 (diff | hist) N User:DrJones1935/29 June 2012 ‎ (Created page with "====I. Check culture and plate for overnight growth==== *''Results: No growth in the ECNR2 culture at 37 oC, confirming that these cultures will not grow at these tem...") (top)
23:13, 3 October 2012 (diff | hist) N User:DrJones1935/28 June 2012 ‎ (Created page with "====I: Inoculate ECNR2 Cultures from Natalie Ma's Plate(from 7 Jun 2012)==== *Inoculate one isolated colony into ~2 mL of LB media in sterile culture tube (x2) *P...") (top)
23:12, 3 October 2012 (diff | hist) Nm User:DrJones1935/27 June 2012 ‎ (Created page with "====I: Make 15% Glycerol Stocks (3 total) from Successful Overnight Culture==== *Transfer 600 uL of each liquid culture to each microcentrifuge tube (x3) :*SIMD43 *Add 200 uL of...") (top)
23:12, 3 October 2012 (diff | hist) N User:DrJones1935/26 June 2012 ‎ (Created page with "====I. Inoculate Liquid Cultures for Cultures Which Failed to Grow==== *Inoculate one isolated colony (or as close as possible) into ~2 mL of appropriate media according to the ...") (top)
23:11, 3 October 2012 (diff | hist) N User:DrJones1935/25 June 2012 ‎ (Created page with "====I. Inoculate Liquid Cultures for Cultures Which Failed to Grow==== *Inoculate one isolated colony (or as close as possible) into ~2 mL of appropriate media according to the ...") (top)
23:11, 3 October 2012 (diff | hist) N User:DrJones1935/22 June 2012 ‎ (Created page with "====I: Make 15% Glycerol Stocks (15 total) from Successful Overnight Cultures==== *Transfer 600 uL of each liquid culture to each microcentrifuge tube (x3 per culture) :*SIMD40 ...") (top)
23:11, 3 October 2012 (diff | hist) N User:DrJones1935/21 June 2012 ‎ (Created page with "====I: Make 15% Glycerol Stocks (42 total) from Successful Overnight Cultures==== *Chill an aliquot of sterile 60% glycerol on ice *Transfer 600 uL of each liquid culture to eac...") (top)
23:03, 3 October 2012 (diff | hist) N User:DrJones1935/20 June 2012 ‎ (Created page with "====I. Make 100 mg/ mL Ampicillin Stock Solution (small) for Plates/Liquid Cultures==== *Weigh out ''(150 mg)'' 150.32 mg of <span style="colo...") (top)
23:02, 3 October 2012 (diff | hist) N User:DrJones1935/15 June 2012 ‎ (Created page with "''Note: Professor Isaacs brought samples of'' B. subtilis ''1A40 from the Breaker Lab. There was 1 plate and 1 liquid culture'' ====I: Make 15% Glycerol Stocks (4) from ''B. sub...") (top)
23:02, 3 October 2012 (diff | hist) N User:DrJones1935/12 July 2012 ‎ (Created page with "====I: Check WT B. subtilis Liquid Culture for Growth==== *RESULTS: Culture appears uniformly cloudy consistent with overnight growth. The glycerol stocks are viable.")

(Latest | Earliest) View (newer 50 | older 50) (20 | 50 | 100 | 250 | 500)