Inverse polymerase chain reaction

Inverse polymerase chain reaction

Inverse polymerase chain reaction (Inverse PCR) is a variant of the polymerase chain reaction that is used to amplify DNA with only one known sequence. One limitation of conventional PCR is that it requires primers complementary to both termini of the target DNA, but this method allows PCR to be carried out even if only one sequence is available from which primers may be designed.

Inverse PCR is especially useful for the determination of insert locations. For example, various retroviruses and transposons randomly integrate into genomic DNA. To identify the sites where they have entered, the known, "internal" viral or transposon sequences can be used to design primers that will amplify a small portion of the flanking, "external" genomic DNA. The amplified product can then be sequenced and compared with DNA databases to locate the sequence which has been disrupted.

The inverse PCR method involves a series of restriction digests and ligation, resulting in a looped fragment that can be primed for PCR from a single section of known sequence. Then, like other polymerase chain reaction processes, the DNA is amplified by the temperature-sensitive DNA polymerase:

# A target region with a internal section of known sequence and unknown flanking regions is identified
# Genomic DNA is digested into fragments of a few kilobases by a usually low-moderate frequency (6-8 base) cutting restriction enzyme.
# Under low DNA concentrations, self-ligation is induced to give a circular DNA product.
# PCR is carried out as usual, with primers complementary to sections of the known internal sequence.


Wikimedia Foundation. 2010.

Look at other dictionaries:

  • Polymerase chain reaction — PCR redirects here. For other uses, see PCR (disambiguation). A strip of eight PCR tubes, each containing a 100 μl reaction mixture The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a single or a… …   Wikipedia

  • Polymerase Chain Reaction — Réaction en chaîne par polymérase Pour les articles homonymes, voir PCR. Diagramme des 4 premiers cycles de la PCR La réaction en chaîne par polymérase (PCR est l abréviation an …   Wikipédia en Français

  • Polymerase Chain Reaction — Die Polymerase Kettenreaktion (englisch Polymerase Chain Reaction, PCR) ist eine Methode, um die Erbsubstanz DNA in vitro zu vervielfältigen. Dazu wird ein Enzym verwendet, die DNA Polymerase. Der Begriff Kettenreaktion beschreibt in diesem… …   Deutsch Wikipedia

  • Nested polymerase chain reaction — A diagram illustrating the method of nested PCR. Nested polymerase chain reaction is a modification of polymerase chain reaction intended to reduce the contamination in products due to the amplification of unexpected primer binding sites.… …   Wikipedia

  • Multiplex polymerase chain reaction — (Multiplex PCR) is a modification of polymerase chain reaction in order to rapidly detect deletions or duplications in a large gene. This process amplifies genomic DNA samples using multiple primers and a temperature mediated DNA polymerase in a… …   Wikipedia

  • Overlap extension polymerase chain reaction — This page assumes familiarity with the terms and components used in polymerase chain reaction (PCR) process. The overlap extension polymerase chain reaction (or OE PCR) is a variant of PCR. It is also referred to as Splicing by overlap extension… …   Wikipedia

  • Inverse Polymerase-Kettenreaktion — Ablauf der inversen PCR. Als Inverse Polymerase Kettenreaktion oder kurz Inverse PCR versteht man eine Abwandlung der PCR, mit der man unbekannte Genbereiche amplifizieren kann. Bei der Inversen PCR benötigt man einen relativ kurzen, aber von der …   Deutsch Wikipedia

  • Inverse PCR — Ablauf der inversen PCR. Als Inverse Polymerase Kettenreaktion oder kurz Inverse PCR versteht man eine Abwandlung der PCR, mit der man unbekannte Genbereiche amplifizieren kann. Bei der Inversen PCR benötigt man einen relativ kurzen, aber von der …   Deutsch Wikipedia

  • Inverse Polymerasekettenreaktion — Ablauf der inversen PCR. Als Inverse Polymerase Kettenreaktion oder kurz Inverse PCR versteht man eine Abwandlung der PCR, mit der man unbekannte Genbereiche amplifizieren kann. Bei der Inversen PCR benötigt man einen relativ kurzen, aber von der …   Deutsch Wikipedia

  • Reaction En Chaine Par Polymerase — Réaction en chaîne par polymérase Pour les articles homonymes, voir PCR. Diagramme des 4 premiers cycles de la PCR La réaction en chaîne par polymérase (PCR est l abréviation an …   Wikipédia en Français

Share the article and excerpts

Direct link
Do a right-click on the link above
and select “Copy Link”