Improved cloning efficiency of polymerase chain reaction (PCR) products after proteinase K digestion

Improved cloning efficiency of polymerase chain reaction (PCR) products after proteinase K digestion

We and others have occasionally experienced difficulty in cloning PCR derived DNA fragments (1). This has occurred even after extraction of the DNA with phenol and phenol:chloroform, followed by precipitation with ethanol and/or column purification of the bands. To test the hypothesis that the Taq polymerase remains bound to the DNA and therefore inhibits restriction endonuclease activity, we i...

Polymerase Chain Reaction (PCR)

The polymerase chain reaction (PCR) is a powerful and widely used technique that has greatly advanced our ability to analyze genes. Genomic deoxyribonucleic acid (DNA) present in cells contains many thousands of genes. This makes it difficult to isolate and analyze any individual gene. PCR allows specific DNA sequences, usually corresponding to genes or parts of genes, to be copied from genomic...

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*

Increasing detection of polymerase chain reaction (PCR) by isolation of PCR products (IPCRp).

AIM To develop a method for enhanced polymerase chain reaction (PCR) product detection. METHODS During the PCR, the double-stranded product is generated with fluorescent dye on one strand, and biotin on the other strand. The product is captured on the streptavidin-coated plates with high efficiency (IPCRp). Washing of the all unamplified compounds, including dye-labeled unincorporated primers...

Development Application of Polymerase Chain Reaction (PCR)

Polymerase chain reaction (PCR) is an in vitro technique to synthesize large quantities of a given DNA molecule that separates the DNA into two complementary strands, uses a peltier heat pump to quickly heat and cool the DNA and uses the Taq polymerase for the synthesis of DNA. Taq is a bacterium that lives by volcanic sulfer jets at the bottom of the ocean where the temperature is very high. F...