Ligation
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'''Quick Ligation:''' | '''Quick Ligation:''' | ||
Latest revision as of 16:46, 8 June 2006
This is the procedure we tried for ligation of hixR, hixL, hixC:
Quick Ligation:
Step 1: Combine 50 ng of vector with a 3-fold molar excess of insert; adjust volume to 10 ul w/ dH2O.
Step 2: Add 10 ul of 2x Quick Ligation Reaction buffer/mix.
Step 3: Add 1 uL of Quick T4 DNA Ligase/mix .
Step 4: Centrifuge briefly/incubate @ (25 C) for 5 minutes.
Step 5: Chill on ice, then transform or store @ -20 C.
