Bio-Rad Ligation and Transformation Module - 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
❒
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