Talk:20.109(S10):Start-up protein engineering (Day1): Difference between revisions

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| Orange
| Orange
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|D78K
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|G GCT AGA AAA ATG AAG GAC ACA GAC AGC GAG GAG GAA ATC CG
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|BseRI
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|Glu 83
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| Yellow
| Yellow
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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