Team:Rutgers/Notebook

The Rutgers iGEM team sets out to find a lab.

Lab Found!

Dr. Sagar Khare in the Center for Integrative Proteomics Research offers us laboratory space and supplies to house the Rutgers iGEM team.

First week of July

Lab safety training: Competent cell preparation, Plate preparation, Plate and cell negative control testing, Confirmation of plate antibiotics and negative controls , Acquisition of CRE recombinase protein from Dr. Pal Maliga at Waksman Institute of Microbiology.

Transform the following: mRFP (I13521), GFP (I13522), CFP (I13600), mOrange (E2050), LuxR (C0062), LuxI (C0061), LuxBox (R0063), pSB1C3, CRE (in pET29 vector)

Second week of July

Start overnights of transformations, Minipreps of genes, Digestions of LuxR and LuxBox, Gel Electrophoresis (failed digestion attempt), Start overnights of transformations, Minipreps of genes, Gel Electrophoresis, Ligation of LuxR and LuxBox, Transformation of L1 ( LuxR+LuxBox)

July 15th-19th

Start overnight of L1, Miniprep L1, Review sequencing data, Failed ligation attempt, Start overnights of transformations, Minipreps of genes, Digestions of LuxR and LuxBox, Gel Electrophoresis (failed digestion attempt), Start overnights of transformations, Minipreps of genes, Gel Electrophoresis, Ligation of LuxR and LuxBox, Transformation of L1 ( LuxR+LuxBox)

July 22nd-26th

PCR of mRFP loxp site addition, PCR of GFP loxp site addition, Gel electrophoresis confirmation, Digestion of LuxR and LuxBox

July's last days

Reconsider CRE recombinase project, Quorum sensing degradation design

August!

New plasmid design for LuxR, LuxI, qsdA degradation, gfp. Order from genscript

First week of September

Arrival of genscript plasmids, Transformation for BBa_K1206000, Miniprep BBa_K1206000, Sequence verification

Mid-September

Assay for fluorescence of GFP in project plasmid