Team:Tsinghua/Notebook/4 July 2011

From 2011.igem.org

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Digest pET15b vector with NdeI + BamHI (Takara), 37 centigrade, 3 hours. (Approx. 1ug)
+
==[[pET15b OmpA-GFP construct]]==
 +
  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.
 +
  YX
-
Recycled digested vector, digested GFP fragment, digested OmpA fragment (previous day) via gel-extraction.
+
==[[pet15b OmpA-HIV cleavege site-sh3]]==
-
 
+
  PCR OmpA-HIV site&HIV site-sh3
-
 
+
  overlapping pcr
-
Ligation
+
  gel extraction
-
 
+
-
1ul vector + 10ul GFP + 10ul Takara Solution I
+
-
 
+
-
RT, 1h
+
-
 
+
-
Transform DH5a competent cells, spread on Amp+ LB plates.
+
-
 
+
-
YX
+

Latest revision as of 13:38, 5 September 2011

pET15b OmpA-GFP construct

 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.
 YX

pet15b OmpA-HIV cleavege site-sh3

 PCR OmpA-HIV site&HIV site-sh3
 overlapping pcr
 gel extraction
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