Team:Austin UTexas LASA/Notebook
Notebook
Sept 1 - Sept 9, 2016
Week 1: Got aliquots of HpaBC from lab, grew up parts (Hpab, HpaC, pp2551) and digested them to check lengths.
Oct 3- Oct 7, 2016
Week 1: Did PCR to get HpaB out of the HpaBC lab plasmid, sequence verified ppddc in BTK001 backbone, sequence verified pp2551 and HpaC, made glycerol stocks of sequence verified parts
Oct 10 - Oct 14, 2016
Week 2: Ordered HpaBC operon from IDT, grew up pEM7 [BTK156], did Gibson assembly of HpaBC in YTK001 entry vector and grew it up
Oct 17 - Oct 21, 2016
Week 3: Split parts into different teams, each team working on assembling and sequence verifying individual parts into entry vecotrs[ppddc, HpaBC pEM7, pp2551]
Oct 24 - Oct 28, 2016
Week 4: Grew up and miniprepped HpaBC, and pp2551, but transformation of HpaBC did not work.
Nov 1- Nov 3, 2016
Week 1: Did Gibson assembly, transformed and miniprepped ppddc, pEM7, and HpaBC, found that Gibson assembly for HpaBC did not work
Nov 6 - Nov 10, 2016
Week 2: Redid Gibson assembly for HpaBC, realized we were cloning our parts into YTK001 [plasmid vector]
Nov 13 - Nov 17, 2016
Week 3: Transformed, miniprepped, made glycerol stocks of HpaBC. Did a gibson assembly of ppddc.
Nov 20 - Nov 24, 2016
Week 4: Transformed pp2551, transformed Gibson product of pEM7, transformed and miniprepped ppddc gibson assembly.
Dec 1- Dec 8, 2016
Week 1: pp2551 plates had no colonies, streaked with glycerol stocks of HpaBC, redid transformation of pp2551
Dec 11 - Dec 15, 2016
Week 2: Transformation of HpaBC, ppddc plates had no colonies, redid Gibson assembly for ppddc
Dec 18 - Dec 22, 2016
Week 3: miniprepped and made glycerol stocks of HpaBC, got very low concentrations for the minipreps, looked into origin of the backbone (BTK001). Transformed gibson reaction of ppddc, plate had no colonies.
Dec 25 - Dec 29, 2016
Week 4: Digested ppddc and backbone (BTK001) to see if there was a problem in the ori, ran on a gel, showed no insert.
Jan 1- Jan 5, 2017
Week 1: Used Zymo DNA clean up kit to get more concentrated values to send in for sequencing. Established plan for how to proceed with each part.
Jan 8 - Jan 12, 2017
Week 2: Miniprepped pp2551. Designed primers for ppddc pcr to add golden gate overhangs.
Jan 15 - Jan 19, 2017
Week 3: Sent in HpaBC and pp2551 to sequencing. Started ‘lab trainings’ with highschool students new to the club.
Jan 22 - Jan 26, 2017
Week 4: Received primers for ppddc, and diluted them.
Feb 1- Feb 9, 2017
Week 1: Ran a PCR reaction of ppddc to add golden gate overhangs.
Feb 12 - Feb 16, 2017
Week 2: Received and grew up an aliquot of new backbone (YTK001) with ColE origin because the original backbone was giving us trouble with getting high enough concentrations to sequence verify it.
Feb 19 - Feb 23, 2017
Week 3: Miniprepped new backbone (YTK001). Did a PCR clean-up of ppddc.
Feb 26 - Feb 28, 2017
Week 4: Re-ran PCR reaction and purified ppddc because we were running low.
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