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KMID : 1204320090250040393
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Laboratory Animal Research
2009 Volume.25 No. 4 p.393 ~ p.395
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Improved Polymerase Chain Reaction Method for Genotyping Lepob Mouse Using Mismatched Allele-Specific Primers
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Rhee Sang-Dal
Jung Won-Hoon
Kim Hee-Youn
Cheon Hyae-Gyeong
Sohn Ji-Young
Kho Min-Soo
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Abstract
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The previously reported methods for genotyping Lepob mice have shortcoming of either inconveniency or poor reproducibility. Therefore we improved up these protocols by adding the mismatched base pairs in the allele-specific primers for stable discrimination of single base pair mismatch and amplifying the polymerase chain reaction (PCR) product in one tube. This new simplified genotyping method results in higher reproducibility even with crudely prepared DNA. When the method was used to select heterozygotic breeders to produce obese mice, 256 pairs of the all 287 mating pairs produced obese offspring, providing 90% or greater genotyping efficiency. This method using PCR with mismatched allele specific primers is appropriate and efficient for genotyping a large number of Lepob mutants.
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KEYWORD
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Allele specific PCR, genotyping, Lepob mice
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