IGEM:MIT/2006/Notebook/2006-7-24: Difference between revisions
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==Smell Experiments w/ Isoamyl | ==Smell Experiments w/ Isoamyl Acetate== | ||
*Questions that we want to answer: | *Questions that we want to answer: | ||
*#Does IAA smell like banana? '''Yes''' | *#Does IAA smell like banana? '''Yes''' | ||
| Line 12: | Line 12: | ||
*2.5μL 0.55ng/μL E. coli genome | *2.5μL 0.55ng/μL E. coli genome | ||
*1.25μL each respective primer (25 ng/μL) | *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? --[[User:Austin|Austin]] 12:20, 24 July 2006 (EDT) | |||
==pchB and pchA primers== | |||
*pchB forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''at gaa aac tcc cga aga ctg cac''' - 3' | |||
*pchB reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT T'''at gcg gca ccc cgt gtc tgg cg''' - 3' | |||
*pchA forward: 5' - GTT TCT TCG AAT TCG CGG CCG CTT CTA G'''at gag ccg gct ggc gcc cct gag''' - 3' | |||
*pchA reverse: 5' - GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TAT T'''ag gcg acg ccg cgc tgc aag g''' - 3' | |||
*pchA mutagenesis forward: 5' - cga ccc att gga ccc gct '''a'''ca ggt att cgg tgc ctg - 3' | |||
*pchA mutagenesis reverse: 5' - cag gca ccg aat acc tg'''t''' agc ggg tcc aat ggg tcg - 3' | |||
Latest revision as of 09:37, 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'
- pchA mutagenesis forward: 5' - cga ccc att gga ccc gct aca ggt att cgg tgc ctg - 3'
- pchA mutagenesis reverse: 5' - cag gca ccg aat acc tgt agc ggg tcc aat ggg tcg - 3'
