June 26
Start overnight cultures of T7 polymerase (K145001), mCherry (J06702), Lac Promoter (R0010), double terminator (B0014 and B0015) and Tet Promoter (R0040)
June 27
Miniprep of T7 polymerase (K145001), mCherry (J06702), Lac Promoter (R0010), double terminator (B0014 and B0015) and Tet Promoter (R0040) and submit for sequencing
Transfer 0.5 mL aliquots of BPA and 5 mL aliquots 17a-ethynylestradiol cultures from June 23 to fresh minimal media
Check DDT and nonylphenol cultures, transfer to fresh minimal media if there is growth
Ethanol precipitation of DNA extractions from LA river samples
Streak out cells from fosmid kit
Fosmid kit first gel?
Prepare antibiotic stocks (Ampicillin, Chloramphenicol)
June 28
Send off continued forward sequencing for HER
Observe fosmid gel, conduct shearing and end repair, run size selection gel (if needed)
Start culture of fosmid cells overnight
This Week
PCR of parts for pNT001 (Test plasmid for K123000)
Purification/Gibson assembly for pNT001
Email other teams
PCR of parts for pNT002 and assembly
Use pNT001 and pNT002 to test degradation parts
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