Pcr Purification Kit
Pcr Purification Kit - Related Videos
12:39
Rapid Genotyping of Mouse Tissue Using Sigma's Extract-N-Amp Tissue PCR Kit
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Cited by 4 •
2008
The complete genotyping of a mouse tail sample, including tissue digestion and PCR readout, is done in one and a half hours using Sigma's SYBR Green Extract-N-Amp Tissue PCR kit.
11:00
Genotyping of Plant and Animal Samples without Prior DNA Purification
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Cited by 14 •
2012
The Direct PCR approach presented here facilitates PCR amplification directly from small amounts of unpurified plant and animal tissue.
08:51
Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)
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Cited by 9 •
2016
Here, ribosomal spacer PCR (RS-PCR) is used together with a miniaturized electrophoresis system as a fast and high resolution method for genotyping S. aureus at moderate costs allowing a high throughput.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
14:07
Genome-wide Gene Deletions in Streptococcus sanguinis by High Throughput PCR
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Cited by 9 •
2012
An efficient genome-wide single gene mutation method has been established using Streptococcus sanguinis as a model organism. This method has achieved via high throughput recombinant PCRs and transformations.
FAQs
Chaotropic salts create conditions that promote attachment of amplified DNA to a silica membrane. This selective binding allows the DNA to remain on the membrane while soluble reaction components move away during washing. The separation is central to the method because it distinguishes the desired PCR product from substances that could compromise later genetic analyses.
Membrane washing removes residual primers, free nucleotides, polymerase, salts, and other PCR reaction components. These substances can interfere with downstream procedures if they remain with the amplified DNA. Their removal helps provide a cleaner template, supporting more reliable sequencing, cloning, genotyping, and fragment-analysis results.
Purified DNA is released from the silica membrane into a small volume of buffer or water. Eluting into a limited volume helps improve the concentration of the recovered template, which is useful when downstream genetic analysis requires a sufficiently concentrated DNA sample. The resulting preparation is therefore better suited for consistent analysis than an uncleaned PCR mixture.
Cleanup reduces substances from the amplification reaction that may interfere with downstream analysis, helping produce clearer and more reproducible results. In genetics, this improved template quality supports examination of amplified DNA through sequencing, genotyping, cloning, or fragment analysis. The purification step therefore strengthens the reliability of conclusions drawn from PCR-derived material.
A typical workflow first uses chaotropic salts to promote binding of amplified DNA to a silica membrane. The membrane is then washed to remove primers, nucleotides, polymerase, salts, and other contaminants. Finally, purified DNA is eluted in a small volume of buffer or water, producing a cleaner template for the selected downstream procedure.
Researchers can use PCR purification when amplified DNA will undergo sequencing, cloning, genotyping, or fragment analysis. These applications benefit from removal of residual PCR components because contaminants may interfere with subsequent procedures. Purification is especially relevant when the quality and concentration of the amplified template affect the clarity and reproducibility of the genetic result.