Bio-Rad Ligation and Transformation Module - Manual (Page 31)

6. Cool tube on ice for 2 min.
7. Once cool, centrifuge briefly to
bring contents to the bottom of
the tube and keep tube at room
temperature.
8. Spin down stock tubes of pJet1.2
vector and the T4 DNA ligase to
collect the contents at the bottom
of the tube.
9. Set up a ligation reaction with the
following reagents.
Reagent Amount
Blunting reaction (already in 9.0 µl
microcentrifuge tube)
T4 DNA ligase 0.5 µl
pJet1.2 blunted vector 0.5 µl
Total 10 µl
10. Close the cap and mix well.
Centrifuge briefly to collect the
contents at the bottom of the
tube.
11. Incubate the tube at room
temperature for 5–10 min.
12. Store the ligation at –20°C or if
proceeding directly to
transformation step, store on ice.
27
Ice