IGEM:MIT/2006/Notebook/2006-8-3: Difference between revisions
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==new pchBA primers== | ==new pchBA primers== | ||
pchA reverse: G TTT CTT TAC TAG TAT TAT Tag gcg acg ccg cgc tg | pchA reverse new: G TTT CTT TAC TAG TAT TAT Tag gcg acg ccg cgc tg [melt temp = 65.6, no hairpin] | ||
pchB forward new: GTT TCT ACG AAT TCG CGG CCG CTT CTA Gat gaa aac tcc cga aga ctg cac | |||
pchA mut forward new: gat cga ccc att gga ccc gct '''a'''ca ggt att cgg tgc | |||
pchA mut reverse new: gca ccg aat acc tg'''t''' agc ggg tcc aat ggg tcg atc |
Revision as of 10:18, 3 August 2006
Template Idea
[[../../Template]]
Primer To Do
- Order new pchBA primers
Miniprep/Digest To Do
- Miniprep BSMT.B0015 LC
- Cut BSMT.B0015 with XP
- Miniprep B0030
- Cut B0030 with ES and SP(bkb)
- Miniprep R0040
- Cut R0040 with ES and SP(bkb)
WG Ligations/Transformations To Do
- Gel extract BSMT.B0015XP
- PCR Cleanup B0030SP backbone
- Ligate BSMT.B0015XP and B0030SP backbone-> transform
- Ligate B0030ES with BSMT.B0015XP in A/CEP
- Ligate B0030ES with BSMT.B0015XP in A/TEP
osmY & E0840 Ligations/Transformations To Do
- osmY and A/CEP and A/TEP
- osmY cut with SpeI
- E0840 cut with XP
- Ligation of osmY cut with S and E0840 cut with XP and pSB1AK3 cut with EP
ATF1 Miniprep/Digest To Do
- Miniprep ATF1.B0015 LC
- Cut ATF1.B0015 with XP
- Gel extract some of ATF1.B0015XP
ATF1 Ligation/Transformation To Do
- Ligate ATF1.B0015XP with B0030ES in A/CEP and A/TESP
IAA Digest
- BAT2 with EP, ligate with A/C and A/TEP
- THI3 with SX
- pSB1AK3 with SX
IAA Ligations
- BAT2EP w/ A/C & A/T BKB
- THI3SX w/ psBIAK3SX->A/K plate
new pchBA primers
pchA reverse new: G TTT CTT TAC TAG TAT TAT Tag gcg acg ccg cgc tg [melt temp = 65.6, no hairpin] pchB forward new: GTT TCT ACG AAT TCG CGG CCG CTT CTA Gat gaa aac tcc cga aga ctg cac pchA mut forward new: gat cga ccc att gga ccc gct aca ggt att cgg tgc pchA mut reverse new: gca ccg aat acc tgt agc ggg tcc aat ggg tcg atc