Study on the construction of standard DXS6804 allelic ladder via molecular cloning and its genetic polymorphism in Chinese yunnan pumi populations / 西安交通大学学报(医学版)

Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

A study on the preparation of STR allelic ladder / 中国法医学杂志

Objective To produce the standard allelic ladder by using the cloning technique. Methods After the amplification and separation of the STR alleles, they were purified and then connected with T-vectors directly. The combinants were transfected into the component E. coli DH5? cells follwed by cloning and plasmid purification. The allelic ladder were then produced by re-amplifying the recombinant plasmid DNA. Results The allelic ladder made in this way can be produced in a lager amount and can be stored in a relatively long period. Conclusion The results demonstrated that the standard allelic ladder generated in this way is more practical in forensic scienc application. This technique in useful for preparation of domestic STR kits.

Study on the construction of standard D12S375 allelic ladder and its genetic polymorphism in Chinese Han, Hui and Wei populations / 中国法医学杂志

s:To resolve the problem of the accuracy and standardization of STR PCR typing in forensic science practice,we have designed a new method to produce standard D12S375 allelic ladder.Seven different PCR amplified D12S375 allelic fragments were isolated from the gel,eluted into the distilled water and reamplified by PCR.The purified allelic fragments were then blunt end subcloned individually into the pUC plasmid vectors and transfected into competent E.coli DH5? TM cells.The sequencing results confirmed that the size and the constructure of the inserts were correct.The recombinant plasmids DNA with 7 inserts were then used as templates for reamplification to generate D12S375 standard ladder, with which the genetic polymorphisms of D12S375 locus in Chinese Han population in Chengdu,Hui population in Gansu and Wei population in Xinjiang were studied.

Genetic polymorphism of DXS6799 locus in three minority populations from Yunnan province / 西安交通大学学报(医学版)

Objective To resolve the problem of the accuracy and standardization of short tandem repeat-polymerase chain reaction(STR-PCR) typing in forensic practice,the molecular clone technology has been used in producing the standard allelic ladders of DXS6799 locus.And the standard allelic ladder was applied in studying the Lisu,Pumi and De-ang populations in Yunnan Province,China.Methods Polymorphism of DXS6799 was analyzed by PCR and PAGE.Molecular cloning technology was employed to construct standard DXS6799 allelic ladder used for DXS6799 genotyping.Results The large quantities of standard allelic ladder of the locus were harvested,and the genetic polymorphisms of DXS6799 locus in three populations were studied.Conclusion The method is of high value for forensic DNA typing to construct standard ladders.DXS6799 is robust for genetic research and forensic application.

Construction of DXS8378 allelic ladder and the genetic polymorphism in three Chinese populations / 西安交通大学学报(医学版)

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.