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PET15b OmpA-GFP construct
From 2011.igem.org
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Digest pET15b vector with NdeI + BamHI (Takara), 37 centigrade, 3 hours. (Approx. 1ug) | 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. | Recycled digested vector, digested GFP fragment, digested OmpA fragment (previous day) via gel-extraction. | ||
Revision as of 14:18, 2 September 2011
Team:Tsinghua/Notebook/4_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.
