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		+	[[Team:Tsinghua/Notebook/5_July_2011]]
		+	Digest pET15b vector with NdeI + BamHI (Takara), 37 centigrade, 3 hours. (Approx. 1ug)
		+	Recycled digested vector, digested GFP fragment, digested OmpA fragment (previous day) via gel-extraction.
		+	Ligation
		+	1ul vector + 10ul GFP + 10ul Takara Solution I. RT, 1h
		+	Transform DH5a competent cells, spread on Amp+ LB plates.
		+
		+
	[[Team:Tsinghua/Notebook/5_July_2011]]		[[Team:Tsinghua/Notebook/5_July_2011]]
	Pick 6 colonies from the plates.		Pick 6 colonies from the plates.

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Revision as of 14:18, 2 September 2011

Team:Tsinghua/Notebook/5_July_2011

 Digest pET15b vector with NdeI + BamHI (Takara), 37 centigrade, 3 hours. (Approx. 1ug)
 Recycled digested vector, digested GFP fragment, digested OmpA fragment (previous day) via gel-extraction.
 Ligation
 1ul vector + 10ul GFP + 10ul Takara Solution I. RT, 1h
 Transform DH5a competent cells, spread on Amp+ LB plates.

Team:Tsinghua/Notebook/5_July_2011

 Pick 6 colonies from the plates.
 Inoculate 5ml LB Amp+.
 Culture PCR.
 Dip less than 1 ul bacteria culture (~3hr in 37 centigrade) into a PCR reaction mix with GFP primers.
 All of the clones have the GFP inserts
 Mini-prep the vectors.