[Constructing standard allelic ladders for four short tandem repeat loci and employing them in a population study on Han Nationality of Chengdu in China].
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.


CONCLUSIONS
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.
Deng
Jian-qiang
JQ
Institute of Forensic Medicine, Sichuan University, Chengdu, Sichuan, 610041 PR China.
Ying
Bing-wu
BW
Shi
Mei-sen
MS
Yan
Jing
J
Jia
Zheng-jun
ZJ
Li
Ying-bi
YB
Wu
Jin
J
Zhang
Ji
J
Hou
Yi-ping
YP
D000483:Alleles; D044466:Asian People / Q000235:genetics; D002681:China; D005828:Genetics, Population / Q000379:methods; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic