TITLE

Optimization, verification and detection of SCoT molecular marker system in soybean

AUTHOR(S)
LI Qiang; SU Er-hu; GAO Ju-lin; SUN Ji-ying; YU Xiao-fang; XIE Min
PUB. DATE
October 2013
SOURCE
Chinese Journal of Oil Crop Sciences;2013, Vol. 35 Issue 5, p491
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Start codon targeted polymorphism (SCoT) is a novel gene targeted marker technique. L25 ( 56 ) orthogonal experiment and single factor experiment design were applied to optimize SCoT - PCR system of soybean in five factors as Mg2+, dNTPs, primer, Taq DNA polymerase and template DNA. The results showed that the optimized system was as follows: a total volume of 20 J L including 2.0 mmol · L¹ Mg2+, 0.2 mmol · L¹ dNTPs, 0.250 μmol · L¹ primer, 1.5U Taq polymerase, 30ng DNA template. The most suitable annealing temperature of primers was 50. 4°C. The optimized SCoT - PCR system was tested on eighteen soybeans, and the result was stable and reliable. From the 82 primer combinations tested, 32 were selected with clear band patterns and abundant polymorphism. Amplifications and genetic verification were carried out on Jiyu 75, local black soybean and progenies from Jiyu 75 X local black soybean. The results showed polymorphism bands and common bands from parents and progeny. The system provided a new technology for evaluation of genetic diversity, construction of genetic linkage map and molecular marker assisted breeding for the soybean.
ACCESSION #
97583601

 

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