Student transformation protocol – Bio-Rad Ligation and Transformation Module User Manual
Page 44
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Student Transformation Protocol
Listed are materials and reagents required at the workstations prior to
beginning the transformation activity.
Instructor’s (Common) Workstation
Quantity
(
✔)
Water bath, heating block, or incubator (37°C)
1
❒
Microcentrifuge (refrigerated, if available)
1
❒
Each student team will require the following items to transform bacteria
with one PCR product.
Materials Required at Each Student Station Quantity/Team (
✔)
Ligation reaction from ligation laboratory (on ice)
5 µl
❒
(Optional) Control plasmid 50–200 ng/µl (on ice)
1 µl
❒
1.5 ml microcentrifuge tubes
4
❒
15 ml culture tube containing 1.5 ml of
1
❒
C-growth medium
Transformation reagent A (on ice)
250 µl
❒
Transformation reagent B (on ice)
250 µl
❒
Sterile inoculating loops
2
❒
Fresh LB Amp IPTG agar plate prewarmed to 37°C
1–2
❒
Fresh starter culture plate of
E. coli HB101
1
❒
200 µl adjustable-volume micropipet and tips
1
❒
1,000 µl adjustable-volume micropipet and tips
1
❒
Marking pens
1
❒
Parafilm sealing film
1 strip
❒
(Optional) Vortex mixer
1
❒
(Optional) Vacuum source
1
❒
40