Usualy the restriction fragment length polymorphisms technique is referred to by PCR-RFLP which means you should do a PCR reaction first. Is there any non-PCR based RFLP technique?
Hi Hiba,
Its old school, but if you have enough DNA a straight up Southern blot would avoid any amplification being necessary.
Frank
Frank is right and depending on the number of sequences of interest in the genome Southern blotting may need 3-10 microgrammes of dna template per sample probed
Hi Hiba
Just to add a little background: In PCR-RFLP you amplify some specific fragment of the genome, cut it with a restriction enzyme (usually 4-cutter because the size of the PCR product is small) and run a gel to see the fragments.
As explained in the two answers above, the old fashioned RFLP is based on Southern blot: you cut the genomic DNA (usually one 6-cutter and one 4-cutter REs, depending on the genome size), run a long gel, transfer the DNA to a membrane and hybridize with a marked cloned fragment (nowadays, you could do it with a PCR fragment), the probe. Please, be aware that the probe was not used in its original whole length, but it was cut and marked internally by oligolabelling (Feinberg & Vogelstein, 1984 Anal Biochem).
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