Bio-Rad Ligation and Transformation Module - Manual (Page 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.
26
70°C water bath