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Thanks for the reaction,
My PCR worked already; I have no idea what went wrong . Can you tell me why you should add BSA or DMSO to your PCR. This is the first thing I hear about this.
later I will try to putt my picture again.
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I think you are wright, Somehow there was some contamination but my gel was verry rare.
| Beverly said: | I could not view the picture either. However, I have found smearing during PCR to be contaimination (other template, primers, bacteria, etc) or too much product ran on the gel at once. Also if you change the stain on the gel, that sometimes changes how your product looks.
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Posted By holeung
on 5/9/2005 12:44 PM
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DMSO or glycerol supposedly increases specificity of primer annealing at the expense of affinity. BSA sometimes increases efficiency of the reaction.
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I have got this before, or something like it, most likely is non-specific primer binding.
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| labrat said: | | Unfortunately I couldn't view the picture for the exact problem, but have you tried adding BSA or DMSO to your PCRs? Also, you could run a sample of your DNA before PCR on a gel and have a look at the quality of the DNA. |
. how are these DMSO and BSA are gng to work plz provide the details.as i am also encountering much smearing. tried all new but still not got what is working wrrong. thnks.
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Hi Samm,
would u please say few words about stringency and stepped pcr ? I also want to know how the annealing temperature affects tha yield of pcr?
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Hi there!
I'm also very interested in the detailed reasons for adding DMSO and/or BSA to the mix.
I have to start up a PCR and I'm gathering all the information I can get to make sure my results are good.
Love Esther
Last edited Sep 12, 2011, 7:41 AM by frog-o-rama
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