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Ligation-Mediated PCR (LM-PCR) (CAT#: STEM-MB-0210-WXH)
Introduction
A major requirement of the conventional polymerase chain reaction (PCR) is that sequences at both ends of the region to be amplified be known. Ligation-mediated PCR bypasses this by requiring knowledge of only one end initially and then adding the second end by ligation of a unique DNA linker.
Principle
Ligation-mediated PCR utilizes small DNA fragments called ‘linkers’ (or adaptors) that are initially ligated to fragments of the target DNA. PCR primers designed to bind to the linker sequences are then used to amplify the target fragments.
Applications
• Determination of primary DNA nucleotide sequences.
• Determination of cytosine methylation patterns .
• Determination of DNA lesion formation and repair.
• Determination of in vivo protein–DNA footprints.
• DNA sequencing, genome walking, and DNA footprinting.
• Determination of cytosine methylation patterns .
• Determination of DNA lesion formation and repair.
• Determination of in vivo protein–DNA footprints.
• DNA sequencing, genome walking, and DNA footprinting.
Procedure
1.Restriction enzyme digestion of the genomic DNA and ligation of specific adapters
2.PCR of restriction fragments using specific primers designed from known sequences and adapter sequences
3.Analysis of the amplified fragments using gel electrophoresis and sequencing
2.PCR of restriction fragments using specific primers designed from known sequences and adapter sequences
3.Analysis of the amplified fragments using gel electrophoresis and sequencing
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