Restriction fragment length polymorphism: Difference between revisions

Revision as of 21:21, 27 June 2014

Revision as of 02:28, 24 July 2014

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== Alternatives ==

The technique for RFLP analysis is, however, slow and cumbersome. It requires a large amount of sample DNA, and the combined process of probe labeling, DNA fragmentation, electrophoresis, blotting, hybridization, washing, and [[autoradiography]] could take up to a month to complete. A limited version of the RFLP method that used [[Oligomer restriction|oligonucleotide probes]] was reported in 1985.<ref name="Saiki1">{{cite journal| url=http://sunsite.berkeley.edu/cgi-bin/ebind2html/pcr/034| title= Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia| last=Saiki| first= RK| coauthorsauthor2= Scharf S, |author3=Faloona F, |author4=Mullis KB, |author5=Erlich HA, |author6=Arnheim N | journal=[[Science (journal)|Science]]| date= Dec 20, 1985| volume=230| issue=4732| pages=1350–1354| doi= 10.1126/science.2999980| pmid= 2999980}}</ref> Fortunately, the results of the [[Human Genome Project]] have largely replaced the need for RFLP mapping, and the identification of many [[single-nucleotide polymorphism]]s (SNPs) in that project (as well as the direct identification of many disease genes and mutations) has replaced the need for RFLP disease linkage analysis (see [[SNP genotyping]]). The analysis of VNTR alleles continues, but is now usually performed by [[polymerase chain reaction]] (PCR) methods. For example, the standard [[National DNA database|protocols]] for [[DNA fingerprinting]] involve PCR analysis of [[CODIS|panels]] of more than a dozen VNTRs.

RFLP is still a technique used in marker assisted selection. Terminal restriction fragment length polymorphism (TRFLP or sometimes T-RFLP) is a [[molecular biology]] technique initially developed for characterizing bacterial communities in mixed-species samples. The technique has also been applied to other groups including soil fungi.