Bio-Rad Ligation and Transformation Module - Manual (Page 44)

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