Bio-rad Ligation and Transformation Module Manual do Utilizador descarregar pdf (Página 30)

Ligation – Quick Guide

1. Label a microcentrifuge tube with

your initials, plant name, and

"ligation."

2. Briefly spin down the stock tubes

of 2x reaction buffer and proof

reading polymerase to collect the

contents at the bottom of the

tube.

3. Set up blunting reaction with the

following reagents.

Reagent Amount

2x ligation reaction buffer 5.0 µl

Purified PCR product 1.0 µl

Sterile water 2.5 µl

Proofreading polymerase 0.5 µl

Total 9 µl

4. Close the cap and mix well.

Centrifuge briefly to collect the

contents at the bottom of the

tube.

5. Incubate the tube at 70°C for

5 min.

70°C water bath

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Bio-rad Ligation and Transformation Module Manual do Utilizador Página 30