Bio-Rad Ligation and Transformation Module User Manual

Page 8

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When

Activity to Complete

Duration

At least 1 day prior to

Run a PCR reaction in thermal

3–4 h

starting the Ligation

cycler to amplify a

and Transformation

gene of interest

module

Electrophorese the PCR

1 h

products (optional)

Purify PCR products

0.5 h

At least 3 days prior to

Prepare LB and LB Amp

0.5 h

the transformation step

IPTG agar plates

At least 2 days prior to

Prepare LB broth

0.5 h

the transformation step

Streak

E. coli on a starter LB

5 min

agar plate

Grow

E. coli starter plate at 37°C

16+ h

As late as possible the

Inoculate starter culture

5 min

day before the

Incubate starter culture at

transformation step

37°C in a shaking

8+ h

water bath or incubator

Day of ligation step

Ligate PCR product

1 h

Note: Bolded steps use reagents from the Ligation and Transformation

module.

4

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