Ligation

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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.