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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 ...
- Polymerase chain reaction - Wikipedia
The polymerase chain reaction ( PCR) is a method widely used...
- Reverse transcription polymerase chain reaction - Wikipedia
RT-PCR. Reverse transcription polymerase chain reaction (...
- Polymerase chain reaction - Wikipedia
Feb 1, 2019 · The standard polymerase chain reaction (PCR) is used to amplify a segment of DNA that lies between two inward-pointing primers. In contrast, inverse PCR (also known as inverted or inside-out PCR) is used to amplify DNA sequences that flank one end of a known DNA sequence and for which no primers are available. Inverse PCR DNA involves digestion ...
- Michael R. Green, Joseph Sambrook
- 2019
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What is inverse PCR?
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In inverse PCR ( Nolta et al., 1996 ), a “restriction-digested” DNA template containing the junction between vector LTR and genomic host DNA, is circularized by self-ligation. The circular-ligated DNA serves as template for inverse PCR since it contains a fragment of known vector LTR sequence. This fragment enables the design of two primers.
Schematic depiction of multiplexed inverse PCR using degenerate primers with a 5′ NNN at the mutation site. 1. Make a master mix consisting of 1 μL of 50 ng/μL template plasmid:19 μL of H 2 O:25 μL of Phusion High-Fidelity PCR Master Mix with HF Buffer (NEB) for each reaction. Add 45 μL of the master mix to each well of a 96-well PCR plate.
The Polymerase Chain Reaction (PCR) with its multiple applications in molecular genetic analysis is the cornerstone of modern basic and applied biomedical research. This chapter focuses on the inverse PCR technique that has been used widely over the last two decades in genotyping and chromosome walking applications for the isolation of unknown ...
