IGEM:Peking/2007/Switch-Notebook/2007-8-14: Difference between revisions
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==Colony PCR test of T1T2-(Tet F-lacZ-pLX010)== | ==Colony PCR test of T1T2-(Tet F-lacZ-pLX010)== | ||
22 colonies are selected for incubation. | |||
==PCR and double digestion of RD1== | |||
===PCR of RD1=== | |||
Retract through electropheresis. | |||
===Double digestion of RD1=== | |||
<pre> | |||
10uL 10*H buffer | |||
2uL SpeI | |||
2uL XhoI | |||
15uL RD1 | |||
71uL ddH2O | |||
------------------- | |||
100uL total | |||
</pre> | |||
Latest revision as of 23:14, 17 August 2007
Ligation of SulA, Rec, SD1-3, SS1-2 with GFP-pLX007
Precipitate for retraction
SulA, Rec, SD1-3, SS1-2, GFP-pLX007 (^^ with XbaI & XhoI)
Ligate with GFP-pLX007
SulA, Rec, SD1-3, SS1-2
Transformed in the evening
Colony PCR test of Prec, SD2-3, SS1-3-(SDY-EGFP-pLX007)
Number of colonies grew up:
No. ---------- Prec 0 SD2 0 SD3 1 SS1 >50 SS2 2 SS3 14
Only SS1 is right. 4 colonies are select for incubation.
Colony PCR test of T1T2-(Tet F-lacZ-pLX010)
22 colonies are selected for incubation.
PCR and double digestion of RD1
PCR of RD1
Retract through electropheresis.
Double digestion of RD1
10uL 10*H buffer 2uL SpeI 2uL XhoI 15uL RD1 71uL ddH2O ------------------- 100uL total
