The ladder is running ok in the same gel. Is it a problem of loading dye? Or something wrong in the gel (but the ladder spread normally). I have previously measured the concentration and purity of template DNA by nanodrop and it was ok so no contamination. I use PCR kits from GeneOn company (Germany).
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1. Ladder is separating fine means there is know problem with the gel.
2. It depends on your PCR product size also, let me know which primers you are using and and what is your expected size.
3. Men while upload the gel photo so that i can give you more suggestions..
good luck...
Is there a problem with your gel loading buffer or did you use the same one for your ladder?
may be the PCR didn´t work and your looking whole DNA? in this case the whole DNA remains in the well because it´s size.
Thanks colleagues for your help I actually got it. Problem causes were:
1- The presence of DNA binding proteins in the sample, such as ligases, phosphatases or
2- Annealing of DNA molecules via long cohesive ends.
To eliminate these effects,
I used 6X DNA Loading Dye which is supplemented with 1% SDS. I heated these samples with SDS at 65°C for 10 min, chill on ice, spin down and load.
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