mRNA purification - (Jun/29/2008 )

hi all..............is mRNA purification is necessary during RT-PCR? Is its avoidable?

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Well, it depends on what your experiment is. If you are going in for cDNA library construction then mRNA purification is a must. If you are only going to amplify a specific gene by single step gene specific RT-PCR, then you can use total RNA; but you have to be sure that there is no contaminating DNA. Hope that helps...

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I did purification of ti\otal RNA using mRNA purification of Bangalore genei.During isolation i got good quality RNA also.But after purification (According to the instruction in the kit ) nothing is seen in the gel...................really i wondered and idont know wht to do??????///// please help me wht to do.i dont know whether the fault is mine during purification or something else...........................

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