how to pick clones for a pcr
TRANSCRIPT
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TIPS
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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.
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You need :
A box of sterile tips
A "lid" box of tips
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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 :
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A human can alsobe helpfull
You need :
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Pick a well isolated colony with a tip
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No matter if you stick the point into the agar
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Scratch the tip with bacteria on the Petri dish to a definite place ...
Little finger up !It’s always hepfull
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... 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.
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Continue this process until you have filled the entire plate of PCR or reach the number of clones to analyze
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Then, carefully removethe lid.
The tips will be retained and will beremoved simultaneously from the PCR plate.
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Discard the lid andput the PCR plate ina thermocycler foramplification
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Music by Wasaru
Pictures from Cellectis bioresearch
Tip from Genome Engineering
The next one by you ?