Student ligation protocol – Bio-Rad Ligation and Transformation Module User Manual
Page 40
Student Ligation Protocol
Note: Before use, the appropriate reagents must be defrosted, thoroughly
mixed, and centrifuged to collect contents at the bottom of the tubes. Refer
to Tasks to Perform Prior to the Ligation Laboratory on page 33 for
details.
Listed below are materials and reagents required at the workstations prior
to starting the ligation activity.
Instructor’s (Common) Station
Quantity
(
✔)
Ice bucket containing stock tubes of:
o 2x ligation reaction buffer
100 µl
❒
o Proofreading polymerase
10 µl
❒
o T4 DNA ligase
10 µl
❒
o pJet1.2 blunted vector
10 µl
❒
o Sterile water
1 ml
❒
Water bath, heating block, or incubator at 70°C
1
❒
Microcentrifuge (refrigerated, if available)
1
❒
Materials Required at Each Student Station
Quantity
(
✔)
Purified PCR product
1–2 µl
❒
1.5 ml microcentrifuge tube
1
❒
10 µl adjustable-volume micropipet and tips
1
❒
Ice bucket
1
❒
Marking pen
1
❒
36