Method for enhancing high GC content DNA fragment amplification efficiency by organic solvent

• Main Authors: MENG LIANGYU, ZHANG ZHIZHOU, YANG XIA, LIU FANG
• Format: Patent
• Language: Chinese; English
• Published: 11.03.2009
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; SPIRITS; VINEGAR; WINE

The invention relates to a method for strengthening the amplification efficiency of DNA segments with high GC content through an organic solvent, which comprises the following steps: firstly, extraction of DNA to be amplified; secondly, configuration and optimization of a PCR system; thirdly, setting and operation of PCR conditions; and fourthly, electrophoretic detection of PCR products, wherein the optimization of the PCR system is to add the organic solvent into the PCR reaction system; and the electrophoretic detection of the PCR products is to adopt 1 percent of sepharose gel, to dye by an EB liquid after glue leaking, to use an ultraviolet light-splitting transmissometer to observe the position of amplified segments in the gel, and to compare the amplified segments with corresponding molecular weight standards. The method has obvious effect of amplifying the DNA segments which are rich in GC alkaline base, so that the amplification of sequences with high GC content is more efficiently and has more purpo