AI Article Synopsis
- The study aims to improve the accuracy and standardization of short tandem repeats (STR) typing by creating standard allelic ladders for four specific loci using molecular cloning technology.
- Researchers utilized PCR to generate allelic fragments, which were then cloned and sequenced, confirming the correct size and construction before creating standardized ladders.
- The produced ladders were used to analyze genetic variation in the Han population of Chengdu, indicating that D1S1676 and D2S2735 loci are reliable for forensic analysis, while D11S1977 and D22S444 loci have limited usefulness.
Article Abstract
Objective: To solve the problems in the accuracy and standardization of short tandem repeats-polymerase chain reaction (STR-PCR) typing, the authors adopted the molecular clone technology in producing the standard allelic ladders of D1S1676, D2S2735, D11S1977 and D22S444 loci and applied them in a population study on the Hans in Chengdu, China.
Methods: PCR was used to produce several different allelic fragments of these loci. PCR products were eluted from the gel and re-amplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pGEMR-T plasmid vectors and the recombinant were transfected into competent E.coli DH5alpha TM cells. The results of sequencing confirmed that the size and the construction of the inserts were correct. The recombinant plasmids DNA with the inserts were then used as template for re-amplification to generate the four loci standard ladders.
Results: The authors succeeded in producing large quantity of standard allelic ladder of these four loci, with which the genetic polymorphisms of these loci in Chengdu Han population of China were studied.
Conclusion: This method is of high value for forensic DNA typing to construct standard ladders. D1S1676, D2S2735 loci are robust for forensic analysis in Chinese Han population, whereas the value of D11S1977 and D22S444 loci is limited.
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