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RAPD PCR


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#1 Palitha

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Posted 16 November 2017 - 07:49 PM

 We have performed RAPD testing using a published protocol. The assay worked well previously the consistent results. However, the results have become inconsistent recently with different band pattern for the control organisms.

 

Previous pattern 

Previous pattern.JPG

 

Current pattern

 

Current pattern.jpg

 

 

We have checked for the working condition of Taq polymerase, dNTP & MgCl2,

We have use re diluted the primers from the stocks.

 

Any suggestions are welcome.


Edited by Palitha, 16 November 2017 - 08:09 PM.


#2 hobglobin

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Posted 17 November 2017 - 01:29 PM

Aren't RAPDs already "shot"? wink.png  Hedrick, P. 1992. Shooting the RAPDs. Nature 355:679–680.

Too many issues can affect the results i.e. banding patterns i.e. any little contamination with other DNA, and other interferences up to gel problems...

I'd try a different method if possible.


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#3 GradMicro

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Posted 19 November 2017 - 11:45 PM

Aren't RAPDs already "shot"? wink.png  Hedrick, P. 1992. Shooting the RAPDs. Nature 355:679–680.

Too many issues can affect the results i.e. banding patterns i.e. any little contamination with other DNA, and other interferences up to gel problems...

I'd try a different method if possible.

True, but in resource limited settings, this is like the only feasible method to try out! 






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