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

TR lab

Team colour	Mutation X#Z	Primer (forward)	Silent mutation	New restriction site
Teaching example	S101L	GG AAC GGC TAC ATC CTC GCT GCG CAG TTA CGT CAC G	Ala 103	FspI
Teaching experiment	M124S	GAT GAA GAA GTT GAT GAA AGT ATA CGG GAA GCA GAT ATC GAT GG	Arg 126	AccI
Red	D20R	GAA GCT TTC TCA TTA TTC CGT AAG GAT GGG GAC GGC ACC ATC [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 example	S101L	GG AAC GGC TAC ATC CTC GCT GCG CAG TTA CGT CAC G	Ala 103	FspI
Teaching experiment	M124S	GAT GAA GAA GTT GAT GAA AGT ATA CGG GAA GCA GAT ATC GAT GG	Arg 126	AccI
Red	E84R	G GAC ACA GAC AGC GAG GAG AAA ATC CGA GAA GCA TTC CGT G	Glu 82	BseR1
Orange	D78K	G GCT AGA AAA ATG AAG GAC ACA GAC AGC GAG GAG GAA ATC CG	Glu 83	BseRI
Yellow	E69K [Agi notes: actually E67K]	GGC AAT GGT ACG ATT TAC TTT CCT AAA TTT CTT ACT ATG ATG GC	Gly63	RSAI
Green	D22H	CA TTA TTC GAC AAG CAT GGA GAC GGC ACC ATC	Gly 23	BsmBI
Blue	Q104R	(5’) GC TAC ATC AGC GCT GCT CGG TTA CGA CAC GTC ATG ACA AAC CTC	Arg 106	AflIII
Pink	E84G	5' G GAC ACA GAC AGC GAG GAG GGA ATC CGA GAA GCA TTC CG	Glu 82	BseRI
Purple	D56K	5’ CAG GAT ATG ATA AAT GAA GTC AAG GCT GAT GGC AAT GGA ACG 3’	Ile52	BclI