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Richard Lab:Amplified insert assembly: Difference between revisions
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Richard Lab:Amplified insert assembly (view source)
Revision as of 17:21, 16 February 2011
, 16 February 2011→Procedure
m (Richard Lab:New Standard Assembly moved to Richard Lab:Amplified Insert Assembly: This is the new name for the protocol) |
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#Add 1μL Antarctic Phosphatase and 6μL AP Buffer to the "vector" digest and incubate until the "insert" digest is done. | #Add 1μL Antarctic Phosphatase and 6μL AP Buffer to the "vector" digest and incubate until the "insert" digest is done. | ||
#Kill all reactions by incubating for 20 mins at 80°C. | #Kill all reactions by incubating for 20 mins at 80°C. | ||
#[[Richard Lab:Ligation|Ligate]] ( | #[[Richard Lab:Ligation|Ligate]] at a molar ratio of 4:1 (insert:vector). | ||
#[[Richard Lab:Electroporation of E. coli|Transform]]. | #[[Richard Lab:Electroporation of E. coli|Transform]]. | ||
#Plate on plates with the same antibiotic as the "vector" resistance. | #Plate on plates with the same antibiotic as the "vector" resistance. | ||
