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Problems with PCR Purification


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4 replies to this topic

#1 lis

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Posted 26 January 2002 - 06:07 AM

I have been trying to purify PCR products with sizes ranging from 150-250bp, to be used for sequencing and cloning. I've been using the Qiagen QIAquick PCR Purification Kit, but everytime I lose nearly all my DNA. Can anyone suggest a remedy?

#2 long

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Posted 26 January 2002 - 09:39 AM

I have used Qiagen PCR kit and Gel purification kit for a while and never have this problem. If my pcr gives me sharp band w/o primer dimer, I would use pcr kit, if not, then gel kit. I usually use 20-30 ul of EB buffer for the final elution, which will give me higher concentraction of DAN.



#3 sidhu

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Posted 27 January 2002 - 04:44 AM

Use qiagen pcr kit, but elute with 10-15ul, if you have smaller fragments, it will works fine.
good luck

#4 sankar

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Posted 01 February 2002 - 10:09 PM

Why not you try and use the eppendorf 5 prime Perfectprep PCR gel cleanup. I works really well

#5 sankar

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Posted 01 February 2002 - 10:11 PM

Why not you try the eppendorf 5 prime Perfectprep gel cleanup kit. It works really well




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