Bio-rad Ligation and Transformation Module Manual do Utilizador descarregar pdf (Página 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 ❒

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Bio-rad Ligation and Transformation Module Manual do Utilizador Página 44