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primers deconamination???


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6 replies to this topic

#1 Bilal Malik

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Posted 30 March 2010 - 08:07 PM

hi everybdy......is their anyway of how to decontaminate the primers.... the primers are contaminated with the DNA for which they were specific.. :).....so any possibility of decontaminating them???/

#2 phage434

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Posted 30 March 2010 - 08:41 PM

No. You get to order them again. Next time aliquot so that contamination has little effect.

#3 gyma

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Posted 31 March 2010 - 01:16 AM

never had this problem before, but i ever ran electrophoresis of some several-year-old primers in a 5%(?) gel and for most of them, i saw a clear strong band. however, primers which had showed weak bands seems also ok, no degradation.
maybe it is the best way to reorder it. but if you have time, you can consider to purify it.

#4 Bilal Malik

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Posted 31 March 2010 - 02:25 AM

but how to purify it....????? from the gel????
well iam reodering them...and it'll gona take some time...so just wanted to knw if there was any way to purify them so that can try it in the mean while...

#5 jamessmith01

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Posted 31 March 2010 - 03:17 AM

Primers are cheap! Where do you buy them from? Ours are from Sigma and usually arrive within two days.

#6 gyma

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Posted 31 March 2010 - 05:48 AM

but how to purify it....????? from the gel????
well iam reodering them...and it'll gona take some time...so just wanted to knw if there was any way to purify them so that can try it in the mean while...

there are commercial kits for small sized dna purification but i think those must be expensive. also you can do the purification without kit, but it will cost extra time for you to learn the method. anyway, i put the link here and hope it will help.
http://www.protocol-...osts/33915.html
by the way, i have never used these methods before. good luck.

#7 phage434

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Posted 31 March 2010 - 05:55 AM

You cannot purify primers from template DNA. No matter how well it works, enough template sneaks through to make use in further PCR reactions impossible.




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