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regading blocking of amplified DNA in the gel wells during gel electrophoresis

regading blocking of amplified DNA in the gel wells during gel electrophoresis - Peptide Forum

regading blocking of amplified DNA in the gel wells during gel electrophoresis - Peptide Forum. Ask and discuss questions on peptide protocols, custom peptide synthesis, peptide identification and peptide sequencing.


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Old 07-13-2009, 04:05 PM
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Default regading blocking of amplified DNA in the gel wells during gel electrophoresis



I am facing a problem in running my amplified DNA in agarose gel. actually it gets struck just at the top near the wells and does not move down, although the size of amplified proct is just 1.2 kb. the ladder run alongwith the samples moves normally. can you please suggest me something to get rid of this problem.
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Old 07-13-2009, 05:41 PM
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Default Re: regading blocking of amplified DNA in the gel wells during gel electrophoresis

Your PCR isn't working.
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Old 07-13-2009, 07:57 PM
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Default Re: regading blocking of amplified DNA in the gel wells during gel electrophoresis

You can:
1. Reduce the amount loaded on gel. best method is to repeat PCR and reduce the DNA template added to the reaction.
2. Clean up the PCR product through a Chromaspin Column (pick appropriate size).
3. Perform RNAse digestion.

Reason is too much nucleic acid--so get rid of RNA or decrease PCR product.
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Old 07-15-2009, 01:22 AM
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Default Re: regading blocking of amplified DNA in the gel wells during gel electrophoresis

Hmm... I know this sounds dumb, but I would like to ask a question. I have been working with PCR and gel electrophoresis but have never thought of this. Why would too much nucleic acid cause blocking of DNA migration into the gel wells? Surely some would migrate still?
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amplified , blocking , dna , electrophoresis , gel , regading , wells


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