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Team:Glendale CC AZ/Notebook/Calendar
From 2013.igem.org
Daily Logs
May 20th
First iGEM meeting of the summer!
1. Developed goals for the summer
2. Reviewed lab techniques
3. Reviewed iGEM website and registry
4. Assigned biobricks of interest and genes of interest to present
May 21st
1. Members began their biobrick presentations.
2. We covered online resources available to the team such as Open Wetware, iGEM Help pages, and Rstudio.
3. Team developed protocol for our Growth Curve Assay.
May 22nd
1. Members continued biobrick and gene presentations.
2. Team split into 5 groups and performed our first Growth Curve Assay.
May 28th
1. Biobrick and gene presentations continued.
2. Reviewed protocol for Polymerase Chain Reaction.
May 29th
1. Biobrick and gene presentations continued.
2. Team covered requirements for designing primers, determined primers for all genes of interest.
June 4th
1. Reviewed protocol for DNA isolation.
2. Biotech student Beau came in to present his project on Deinococcus radiodurans.
June 5th
1. Performed Hydrogen Peroxide Growth Curve Assay to determine appropriate concentration of hydrogen peroxide.
June 6th
1. Performed NaCl Growth Curve Assay to determine appropriate concentration of NaCl.
2. Planned next stages of our project.
June 10th
1. Team presented primer designs.
June 11th
1. Performed Hydrogen Peroxide Growth Curve Assay with different concentrations of hydrogen peroxide.
2. Team searched for homolog genes of interest in D. hopiensis.
June 12th
1. Team double checked primer designs for restriction sites Xba1, Pst1, Ecor1, and Spe1.
June 13th
1. Performed Hydrogen Peroxide Growth Curve Assay, still searching working with different concentrations of hydrogen peroxide.
2. Presentation on iGEM's transformation protocol.
June 14th
1. Team went to ASU for meet-and-greet luncheon with ASU's 2013 iGEM team.
June 17th
1. De-briefed members unable to attend ASU luncheon.
2. Fern and Cristina debuted first clips of our team stop motion video!
June 18th
1. Added prefix and suffix sequences to primers, reviewed entire sequence once more prior to ordering.
2. Presentation on Alkaline Lysis Plasmid Miniprep protocol, drew up flowchart.
June 19th
1. Presentation on Gel Electrophoresis protocol.
June 20th
1. Performed Miniprep and Gel Electrophoresis.
June 25th
1. Performed Miniprep.
June 26th
1. Ran yesterday's miniprep products out on a gel.
2. Performed NaCl Growth Curve Assay.
3. Developed protocol and flowchart for Survival Growth Assay.
June 27th
1. Performed NaCl Growth Curve Assay, working with higher concentration of IPTG.
2. Performed Survival Growth Assay.
July 1st
1. Presentation on adding information and navigating our wiki page.
2. First round of wiki sections assigned.
July 2nd
1. Team split up and performed NaCl Growth Curve Assay on transformed E.coli containing PprI gene or RecA gene (depending on group).
2. Performed Survival Growth Assay.
July 3rd
1. Performed transformation on LacI promoter (BBa_R0010), Ribosome Binding Site (BBa_B0034), Double Terminator (BBa_B0015), and RFP Control provided by iGEM.
July 8th
1. Set up bacterial growth culture of transformation products from 7/8/13.
2. Performed Polymerase Chain Reaction.
July 9th
1. Performed NaCl Growth Curve Assay on E.coli containing PprA and RecA.
2. Ran gels of PCR products.
3. Performed Miniprep.
4. Performed Survival Growth Assay.
July 10th
1. Counted colonies on plates from yesterday's Survival Growth Assay.
2. Performed PCR
3. Performed Miniprep.
July 15th
1. Ran gel of of 7/9/13 and 7/10/13 Miniprep products.
2. Ran gel of PCR products from 7/10/13
3. Presentation on Restriction Digest protocol and created table for 7/16/13 digest.
July 16th
1. Performed Restriction Digest according to iGEM's restriction digest protocol.
2. Reviewed iGEM's ligation protocol and created table for 7/17/13.
July 17th
1. Ran gel of digest products to ensure digest was successful.
2. Performed ligation according to iGEM's ligation protocol.
July 18th
1. Team split up to work on transformation and DNA isolation.
July 19th
1. First round of wiki assignments due.
July 22nd
1. Performed Alkaline Lysis Plasmid Miniprep on half of the transformed bacteria from 7/18/13.
2. Prepared 2x 50 mL agarose gels to run all miniprep samples.
July 23rd
1. Performed Alkaline Lysis Plasmid Miniprep on the rest of the transformed bacteria from 7/18/13.
2. Read the absorbance values for all miniprep samples.
3. Read the absorbance values for genomic DNA isolated on 7/18/13.
4. Performed Restriction Digest on miniprep samples.
July 24th
1. Ran miniprep samples on agarose gels prepared 7/22/13.
July 25th
1. Streaked plates with archived glycerol stocks.
2. Performed PCR.
July 29th
1. Performed Alkaline Lysis Plasmid Miniprep on RFP Control, LacI Promoter, Double Terminator, and Ribosome Binding Site.
2. Read absorbance values of miniprep products.
3. Determined volume of miniprep products needed to perform Restriction Digest.
July 30th
1. Ran large agarose gel of PCR products from 7/25/13.
2. Performed PCR on yesterday's miniprep products.
3. Performed PCR on LEA (D. hopiensis).
4. Ran agarose gel of PCR products containing miniprep products from 7/29/13.
July 31st
1. Performed Restriction Digest according to values determined on 7/29/13.
2. Inoculated bacterial cultures from 7/25/13 plates.
August 1st
1. Performed Restriction Digest.
2. Performed Alkaline Lysis Plasmid Miniprep on double parts.
3. Read absorbance values of miniprep products.
4. Ran agarose gel containing digested parts (from earlier today).
5. Performed PCR on single and double parts (LacI promoter, ribosome binding site, double terminator, RFP, LacI+ RBS, TT+ LEA)
6. Performed PCR on D. hopiensis isolates and LEA.
7. Ran out 2x agarose gels containing (separately) PCR products from today.
August 5th
1. Debriefed team on results from all procedures performed 8/1/13.
2. Performed Ecor1 digest using miniprep products from 8/1/13.
3. Re-ran agarose gel of double digested parts from 8/1/13.
August 6th
1. Ran out agarose gel containing PCR products from 8/1/13 (isolates+LEA).
2. Performed Gel Electrophoresis (ran gel) containing Ecor1 digests, as well as select single/double digested parts from previous trials.
August 7th
1. Performed ligation of single and double digests according to iGEM's ligation protocol.
August 8th
1. Performed transformation of all ligation products from 8/7/13 according to iGEM's transformation protocol.
2. Found a few extra biobricks to order from iGEM for easier assembly.
August 10th
1. Team met at local public library to divide up wiki sections and create a complete outline of project section of the wiki.
August 12th
1. Performed PCR of LEA 1 and LEA 2 D. hopiensis isolates.
August 13th
1. Performed PCR of LEA 1 and LEA 2 D. radiodurans isolates.
2. Performed Gel Electrophoresis containing 8/12/13 PCR products.
3. Performed Gel Electrophoresis containing PCR products from today (LEA 1 & 2- D. radiodurans).
August 14th
1. Discussed and planned future of GCC's 2013 iGEM team.
August 15th
1. Performed control restriction digest containing RFP- control ONLY, but different enzymes and different buffers for each. (To ensure enzymes and buffers are performing as expected).
August 16th
1. Performed Gel Electrophoresis containing digest products from 8/15/13.
August 17th
1. Reviewed what we've learned from gel ran on 8/16/13.
2. Performed Alkaline Lysis Plasmid Miniprep on sample A of single parts(LacI promoter, RBS, LEA, TT, and the RFP control).
3. Read absorbance values of miniprep products.
August 20th
1. Performed Gel Electrophoresis on PCR products (Ran gel too long, had to re-run).
2. Performed Alkaline Lysis Plasmid Miniprep on sample B of single parts (LacI promoter, RBS, LEA, TT, and the RFP control).
3. Prepared 2x 40 mL gels for mini prep products from 8/17/13 and earlier today.
4. Read absorbance values for miniprep samples from today.
5. Performed PCR on miniprep products from 8/17/13 as well as miniprep products from earlier today.
6. Prepared samples for sequencing.
August 22nd
1. Performed PCR on LEA 1 and LEA 2 from D. hopiensis.
2. Inoculated bacteria containing single parts (LacI, RBS, TT, LEA-soybean, and RFP).
