Difference between revisions of "Team:BNU-China/Protocol"

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                    <h3>4) Fusion PCR</h3>
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                    <p>1. basic PCR</br>2. use the PCR product of step 1 as template to do PCR</br>3. use the PCR product of step 2 as template to do PCR, but first five cycles don’t add primer, add primer at the sixth cycle and continue PCR.</p>
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                    <p>The system of step 2:</br>
 +
H2O 21μL</br>
 +
2x primeSTAR 25μL</br>
 +
R+F-Primer 2 μL</br>
 +
Template1 1μL</br>
 +
Template2 1μL
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</p>
 
                      
 
                      
 
                      
 
                      

Revision as of 18:08, 17 October 2018

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Protocol

1. PCR system

1) 20μL system is used for verification

Taq:

Super mix:

2) 50μL system for PCR target genes

Pfu mix:


DNA: plasmid 50ng; genome 100~200ng

3) 20μL PCR system for microbes

Super mix:


Bacteria (after picking on the board, back up on the new board and rinse it in the system that has been prepared)

4) Fusion PCR

1. basic PCR
2. use the PCR product of step 1 as template to do PCR
3. use the PCR product of step 2 as template to do PCR, but first five cycles don’t add primer, add primer at the sixth cycle and continue PCR.

The system of step 2:
H2O 21μL
2x primeSTAR 25μL
R+F-Primer 2 μL
Template1 1μL
Template2 1μL