Team:Caltech/Week 2

June 19

Collection of soil and water samples from LA River

June 20

Lab walkthrough
Setting up lab, stock supplies, preparation of LB-amp plates
Test extraction of DNA from BioBrick wells into cloning plates
Initial enrichment cultures of collected samples on minimal media with BPA or 17a-ethynylestradiol

June 21

MoBio kit extraction of DNA from collected samples
Initial enrichment cultures of samples in minimal media with nonylphenol or DDT
Geneious and primer design tutorial

June 22

Miniprep and sequencing of selected BioBricks (K123000, K123001, K123002, K123003)
Preparation of competent cells

June 23

Transform T7 polymerase (BBa_K145001), mCherry (BBa_J06702), Lac Promoter (BBa_R0010), and double terminator (BBa_B0014 and BBa_B0015) biobricks for test sequences for BisA and BisB degradation genes (K123000 and K123001)
Retest competent cells made June 22nd with K123000 and pUC
Transfer 0.5 mL aliquots of BPA and 17a-ethynylestradiol cultures from June 20 to new tubes of minimal media
Design primers for test sequence of BisdA (pNT001)
Design primers to continue sequencing of human ER (K123003)
We determined that sequences for K123000 and K123001 have correct protein coding but different codons than in the sequences shown online

June 24

Transform Tet repressible promoter BBa_R0040</br> Make plain LB and pour plain LB plates</br> Make SOC media</br> Transfer 0.5 mL DDT and nonylphenol cultures from June 21 to new tubes of minimal media
Wash dishes from last year
Design reverse primer for continued sequencing of human ER (K123003)
Update Wiki with protocols we have been using

June 25

June 26

Start overnight cultures of T7 polymerase (BBa_K145001), mCherry (BBa_J06702), Lac Promoter (BBa_R0010), double terminator (BBa_B0014 and BBa_B0015) and Tet Promoter (BBa_R0040)