Bio-rad Ligation and Transformation Module Manuale Utente Scaricare il pdf (Pagina 8)

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.

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Bio-rad Ligation and Transformation Module Manuale Utente Pagina 8