I’m not sure why my PCR bands aren’t clean and rectangular. I am using a 1% gel (50 mL TAE buffer with 0.75 g agarose), and I am loading 1 uL of 6x loading dye with 5 uL of PCR sample. I run at 120 volts for 30 minutes.
0.75g of agarose for 50ml TAE buffer is a 1.5% gel, for 1% you should be using 0.5g for 50 ml.
Hello Brandon, how are you?
Maybe you should check the quality of the buffers you used for the DNA electrophoresis. Sometimes, when the solution is really old, the pH isn't good enough, and that could affect the electrophoresis; try to check if the temperature of your buffer is increasing while the test is running, and if that happens, perhaps it would be a better idea if you prepare again from 0 your buffers for the gel and the run.
Try it and tell me if it works. Anyway, try using less voltage (100V 25 min). That is how I always run my samples.
Regards,
One problem is that your sample lanes are overloaded. Reduce the amount you load by maybe 3-fold. You may also have a lot of salt or similar in those samples but that might also be diminished with smaller sample loadings.
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