IGEM:MIT/2006/Notebook/2006-7-24: Difference between revisions
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Did we get transformants using the annealed primers? NO! :( | Did we get transformants using the annealed primers? NO! :( | ||
*Did you use T4 PNK to 5'-phosphorylate your primers? --[[User:Austin|Austin]] 12:20, 24 July 2006 (EDT) | |||
==pchB and pchA primers== | ==pchB and pchA primers== | ||
Revision as of 09:20, 24 July 2006
Smell Experiments w/ Isoamyl Acetate
- Questions that we want to answer:
- Does IAA smell like banana? Yes
- Does IAA smell strongly in LB?
- At what IAA concentration can E. coli "normal" growth?
- Culture Top10s and add different concentrations of IAA after cells reach OD ~ 0.5
osmY PCR
Done with each pair of new primers meant for PCRing both the short and long osmY promoter region.
- 45μL PCR SuperMix
- 2.5μL 0.55ng/μL E. coli genome
- 1.25μL each respective primer (25 ng/μL)
Did we get transformants using the annealed primers? NO! :(
- Did you use T4 PNK to 5'-phosphorylate your primers? --Austin 12:20, 24 July 2006 (EDT)
pchB and pchA primers
- pchB forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA Gat gaa aac tcc cga aga ctg cac - 3'
- pchB reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT Tat gcg gca ccc cgt gtc tgg cg - 3'
- pchA forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA Gat gag ccg gct ggc gcc cct gag - 3'
- pchA reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT Tag gcg acg ccg cgc tgc aag g - 3'
