Please find attached herewith the electrophoresis results of pcr assay. smearing is consitent finding.. this is a pic of gradient pcr
The strong signal at the bottom of the gel is primer dimer and means that you are using far too much primer. Try using 1/4 0r 1/8th the amount of primer or use a hot start polymerase. If the 2 small bands on the left side of the gel are the correct size then they tell you which temperature to use and I think that the long smearing is simply that you are using far too much dna and this is degraded and produces the smear and that this is possibly interfering with the pcr cycling. Try running a pcr with non amplified dna (no taq polymerase)(same volume as used in this pcr) and also run a pcr at the temperature that does work at 1/2, 1/4. 1/8 and 1/16 the amount of dna in the pcr assay ( serial dilutions) and you may find that less dna and less primer gives more amplification. If your non amplified dna gives the smear then it suggests that you are using too much dna.
@Paul Rutland thank you for the answer... Will try the points raised by you... Can electrophoresis condition leads to smearing
If the buffer is reused and has bugs growing in it then the dna can be degraded or if the voltage is so high that the gel gets hot and softens then you can get smearing.Your gel does seem to have been run for a long time but the small band above the primer dimer does not seem to be smeared
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