Talk:20.109(S10):Start-up protein engineering (Day1): Difference between revisions
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| Orange | | Orange | ||
| | |D78K | ||
| | |G GCT AGA AAA ATG AAG GAC ACA GAC AGC GAG GAG GAA ATC CG | ||
| | |BseRI | ||
| | |Glu 83 | ||
| Yellow | | Yellow | ||
| | | |
Revision as of 14:44, 10 March 2010
TR lab
Team colour | Mutation X#Z | Primer (forward) | Silent mutation | New restriction site |
Teaching sample | S101L | GG AAC GGC TAC ATC CTC GCT GCG CAG TTA CGT CAC G | Ala 103 | FspI |
Red | D20R | GAA GCU UUC UCA UUA UUC CGU AAG GAU GGG GAC GGC ACC AUC [Agi notes: are we ordering RNA or DNA?] |
Ala15 | HindIII |
Orange | G25P | TTA TTC GAC AAG GAT GGG GAT CCC ACC ATC ACC ACA AAG GAA CTT [Agi ordered: C GAC AAG GAT GGG GAT CCC ACC ATC ACC ACA AAG GAA C Primer did not end in G or C, and Tm still is fine.] |
Asp24 | BamHI |
Yellow | Mutation G21W and G23W [Agi notes: actually G23 and G25] | primer: C GAC AAG GAT TGG GAC TGG ACC ATC ACC ACA AAG GAA CTT GGC ACC G[Agi ordered: GAC AAG GAT TGG GAC TGG ACC ATC ACC ACA AAG GAA CTT GGT ACC. Primer was too long and missing silent change - I only noticed latter because I had to make sure I wasn't chopping it off.] | Silent mutation changes a C to a T at position 302 in the pericam. (position 32 in calmodulin). | Enzyme KpnI |
Green | G23P | CA TTA TTC GAC AAG GAT CCG GAC GGC ACC ATC ACC ACA AAG G | none | BamHI |
Blue | D129P | GTT GAT GAA ATG ATC AGG GAA GCA CCT ATC GAT GGT GAT GGC C | Ile 403 | BclI |
Pink | G60P [Agi notes: actually G59P] | 5' G CAG GAT ATG ATC AAT GAA GTC GAC GCT GAT CCC AAT GGA ACG 3’ | Asp 57 | Acc1 |
Purple | N137F | 5’ - C GAT GGT GAT GGC CAA GTA TTC TAT GAG GAG TTT GTA CAA ATG – 3' | Glu 139 | BseRI |
WF lab
Team colour | Mutation X#Z | Primer (forward) | Silent mutation | New restriction site | |||||
Teaching sample | S101L | GG AAC GGC TAC ATC CTC GCT GCG CAG TTA CGT CAC G | Ala 103 | FspI | |||||
Red | |||||||||
Orange | D78K | G GCT AGA AAA ATG AAG GAC ACA GAC AGC GAG GAG GAA ATC CG | BseRI | Glu 83 | Yellow | ||||
Green | |||||||||
Blue | Q104R | (5’) GC TAC ATC AGC GCT GCT CGG TTA CGA CAC GTC ATG ACA AAC CTC | Arg 106 | AflIII | |||||
Pink | |||||||||
Purple |