TIPS

How to pick clones for a PCR

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You need :

A plate containing the PCR amplification mix

Primers, DNA pol, etc.

It exists on the market various ready-mix, where only the primers (and bacteria) must be added.

You need :

A box of sterile tips

A "lid" box of tips

A petri dishcontaining LB agar + antibiotic selection

Usually amp or kana

This petri dish must be gridded and each square should be numbered.

You need :

A human can alsobe helpfull

You need :

Pick a well isolated colony with a tip

No matter if you stick the point into the agar

Scratch the tip with bacteria on the Petri dish to a definite place ...

Little finger up !It’s always hepfull

... then put the tip to a definite place of the PCR plate

For example,

the location #1 on the Petri dish is the location A1 on the PCR plate,

#2 is A2,

#3 is A3,

etc.

Continue this process until you have filled the entire plate of PCR or reach the number of clones to analyze

Then, carefully removethe lid.

The tips will be retained and will beremoved simultaneously from the PCR plate.

Discard the lid andput the PCR plate ina thermocycler foramplification

Music by Wasaru

Pictures from Cellectis bioresearch

Tip from Genome Engineering

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