Team:FDR-HB Peru/notebook
Notebook
February 28
Objective: Make antibiotic solutions CAM, AMP, and KAN. Make stock solutions for the cells that we might use.
March 14
Objective: Transform RFP.
March 27 - May 2
Objective: Practice plasmid preps, restriction reactions and learn how to best use our GreenGel stain in electrophoresis.
May 26
Objective: our iGEM team met with TASA.
May 29
Objective: Isolate BBa_k1724000 from distribution kit, TFX with the same plasmid.
May 31
Objective: make glycerol stock of BBa_k1724000 transformed bacteria.
June 14
Objective: create a growth curve of bacteria with fishmeal.
August 16-August 22
Objective: make glycerol stock of BBa_k1724000 transformed bacteria.
September 5
Objective: create a growth curve of bacteria with fishmeal.
September 13
Objective: Transform competent cells with K1724000 plasmid.
September 27
Objective: Do a mini prep with MerR, pCadA and RFP plasmids to obtain more DNA. Results: We obtained 25µl of PCadA and 50µl of MerR and RFP each.
October 2
Objective: Run a gel with MerR, pCadA and RFP plasmids to obtain DNA concentrations.
October 4
Objective: Digest MerR, pCadA and RFP plasmids to obtain vector and inserts and check for complete digestion. Results: CadA and RFP were mostly digested, MerR was partly digested.
October 11
Objective: Gel purify pCadA vector and RFP, MerR inserts.
October 12
Objective: Check purified linearized pCadA & RFP and MerR inserts. Results: MerR had two bands so we needed to purify the gel again.
Objective: pCadA + RFP ligation and transformation of ligation products.
October 13
Objective: Inoculate the colonies of the transformation of ligation products . Results: Almost no colonies (a few pink colonies of the 1:4 ratio) due to oven being at 39.9 degrees celsius. Inoculated LB CAM with every colony that we found - some from 1:2, 1:4, and 1:5 ratios (nothing from 1:3).
October 14
Objective: Digest RFP to check if it formed the correct construct. Results: Incorrect construct. The correct construct would yield a 2070 bp vector + 900 bp insert. Instead, we obtained a 2939 bp vector and 31 bp insert and the RFP ligated the wrong way.
October 15
Objective: Plasmid prep - Red 1:4 4ul and Digest - 10ul of both Pcad A & Pcad A + RFP.