Turbot fluorescence labeling microsatellite sextuple PCR family tree recognizing method

• Main Authors: RUAN XIAOHONG, WANG WEIJI
• Format: Patent
• Language: Chinese; English
• Published: 31.08.2011
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention relates to a turbot fluorescence labeling microsatellite sextuple PCR family tree recognizing method which comprises the following steps: using three kinds of fluorescence (6FAM, HEX and NED) for pairwise labeling six pairs of primers according to the sizes of the PCR amplifying products of the primers; using primer design software for excluding the possibility that the six pairs ofprimers form dipolymer in the PCR reaction process, and combining the six pairs of primers in the same PCR reaction system which comprises ROX fluorescence molecule internal labels; adjusting the PCRreaction system, and establishing parameters which can enable the six pairs of primers to be simultaneously amplified in the same PCR reaction system and a proper PCR reaction procedure; and analyzing PCR products in six microsatellite positions of the turbot through ABI3130 or a similar gene analyzer according to the fluorescence labels. The sextuple PCR technology can improve the detection efficiency of the turbot micros