Establishing an accurate and rapid method for copy number and zygosity determination can accelerate genetic engineering research process. In this study, droplet digital PCR (ddPCR), an emerging DNA absolute quantifica...Establishing an accurate and rapid method for copy number and zygosity determination can accelerate genetic engineering research process. In this study, droplet digital PCR (ddPCR), an emerging DNA absolute quantification technology, was used to identify single-copy homozygous transgenic lines from a batch of T0 transgenic rapeseed harboring 11 exogenous elements. Copy number of exogenous gene was evaluated in T0 generation based on calculated ratio between transgene and reference CruA gene, single-copy transformants were selected for selfing followed by subsequent zygosity analysis. Single-copy homozygous transgenic plants were successfully screened out in T1 generation by ddPCR.Segregation analysis with T2 seedlings verified that identification results of ddPCR were accurate and reliable. This study provides a novel rapid and accurate method for copy number and zygosity determination in transgenic rapeseed which overcomes disadvantages of traditional Southern analysis and recently developed real-time quantitative method.展开更多
To investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist ...To investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist Methods Comparative genomic hybridization (CGH) was used to define global chromosomal aberrations in 43 primary gastric tumors Based on the results of CGH, analysis of loss of heterozygosity (LOH) was performed in chromosome 19 in which the loss was first discovered in the gastric cancers The PCR-based approach was used to investigate 22 loci, which are spaced at 1 1-10 9 cM intervals throughout chromosome 19 The amplified PCR fragments were subjected to electrophoresis in PAGE gel and analyzed with Genescan TM and Genotyper TM Results CGH analysis revealed gains in chromosome 3p(8/43), 8q(8/43), 20 [20 (9/43), 20p(7/43), 20q(4/43)], 12q(16/43), 13q(12/43) and losses in 19 [19 (15/43)], 7 [17 (8/43), 17p (10/43)], 16 (10/43) and 1p (11/43) Among the 43 evaluated samples, the most frequent LOH was detected at locus D19S571 (27 81%) Conclusions The tumorigenesis of gastric cancer includes several chromosomal changes The aberration of chromosome 19 was the first common change founded in gastric cancer The region near the D19S571 might harbor potential genes related to the tumorigenesis of gastric cancer展开更多
Characterization of genetically modified organisms through determination of zygosity and transgene integration concerning both copy number and genome site is important for breeding a transgenic line and the use ...Characterization of genetically modified organisms through determination of zygosity and transgene integration concerning both copy number and genome site is important for breeding a transgenic line and the use of these organisms in the purpose for which it was obtained.Southern blot,fluorescence in situ hybridization or mating are demanding and time-consuming techniques traditionally used in the characterization of transgenic organisms and,with the exception of mating,give ambiguous results.With the emergence of the real-time quantitative PCR technology,different applications have been described for the analysis of transgenic organisms by determination of several parameters to transgenic analysis.However,the accuracy in quantitation by this method can be influenced in all steps of analysis.This review focuses on the aspects that influence pre-analytical steps(DNA extraction and DNA quantification methods),quantification strategies and data analysis in quantification of copy number and zygosity in transgenic animals.展开更多
Triplets are seldom and monozygotic triplets are scarcely seen in humanity. There are few reports on triplets at home and abroad and diagnosis of zygosity is rarely made. We did the analysis of clinical material on tw...Triplets are seldom and monozygotic triplets are scarcely seen in humanity. There are few reports on triplets at home and abroad and diagnosis of zygosity is rarely made. We did the analysis of clinical material on two sets of triplets. Diagnosis and analysis of zygosity were made in eleven ways about 57 items including blood type and RBC enzyme type, etc.展开更多
The study of twins provides a unique opportunity to evaluate the genetic and nongenetic contributions to a variance of multitude of traits.Twin research has revealed the impact of a strong genetic component to the var...The study of twins provides a unique opportunity to evaluate the genetic and nongenetic contributions to a variance of multitude of traits.Twin research has revealed the impact of a strong genetic component to the variance of many dental traits.The present study was conducted to assess the dental caries and oral hygiene status in monozygotic(MZ)and dizygotic(DZ)twins.A random sample of 129 MZ and DZ twin pairs aged 6-15 years were included in this study.The zygosity of twin pairs was initially recorded by facial appearance and later determined by dermatoglyphics.Dental caries and oral hygiene status were recorded in these twin pairs and the data were subjected to statistical analysis.No significant difference was observed in the mean decayed,extracted due to caries and filled teeth(deft)in primary dentition and decayed,missing and filled(DMFT)in permanent dentition scores and oral hygiene status in MZ and DZ twin pair&The similarity observed in oral health status reflects the effects of a shared environment in MZ and DZ twin pairs.展开更多
文摘Establishing an accurate and rapid method for copy number and zygosity determination can accelerate genetic engineering research process. In this study, droplet digital PCR (ddPCR), an emerging DNA absolute quantification technology, was used to identify single-copy homozygous transgenic lines from a batch of T0 transgenic rapeseed harboring 11 exogenous elements. Copy number of exogenous gene was evaluated in T0 generation based on calculated ratio between transgene and reference CruA gene, single-copy transformants were selected for selfing followed by subsequent zygosity analysis. Single-copy homozygous transgenic plants were successfully screened out in T1 generation by ddPCR.Segregation analysis with T2 seedlings verified that identification results of ddPCR were accurate and reliable. This study provides a novel rapid and accurate method for copy number and zygosity determination in transgenic rapeseed which overcomes disadvantages of traditional Southern analysis and recently developed real-time quantitative method.
文摘To investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist Methods Comparative genomic hybridization (CGH) was used to define global chromosomal aberrations in 43 primary gastric tumors Based on the results of CGH, analysis of loss of heterozygosity (LOH) was performed in chromosome 19 in which the loss was first discovered in the gastric cancers The PCR-based approach was used to investigate 22 loci, which are spaced at 1 1-10 9 cM intervals throughout chromosome 19 The amplified PCR fragments were subjected to electrophoresis in PAGE gel and analyzed with Genescan TM and Genotyper TM Results CGH analysis revealed gains in chromosome 3p(8/43), 8q(8/43), 20 [20 (9/43), 20p(7/43), 20q(4/43)], 12q(16/43), 13q(12/43) and losses in 19 [19 (15/43)], 7 [17 (8/43), 17p (10/43)], 16 (10/43) and 1p (11/43) Among the 43 evaluated samples, the most frequent LOH was detected at locus D19S571 (27 81%) Conclusions The tumorigenesis of gastric cancer includes several chromosomal changes The aberration of chromosome 19 was the first common change founded in gastric cancer The region near the D19S571 might harbor potential genes related to the tumorigenesis of gastric cancer
文摘Characterization of genetically modified organisms through determination of zygosity and transgene integration concerning both copy number and genome site is important for breeding a transgenic line and the use of these organisms in the purpose for which it was obtained.Southern blot,fluorescence in situ hybridization or mating are demanding and time-consuming techniques traditionally used in the characterization of transgenic organisms and,with the exception of mating,give ambiguous results.With the emergence of the real-time quantitative PCR technology,different applications have been described for the analysis of transgenic organisms by determination of several parameters to transgenic analysis.However,the accuracy in quantitation by this method can be influenced in all steps of analysis.This review focuses on the aspects that influence pre-analytical steps(DNA extraction and DNA quantification methods),quantification strategies and data analysis in quantification of copy number and zygosity in transgenic animals.
文摘Triplets are seldom and monozygotic triplets are scarcely seen in humanity. There are few reports on triplets at home and abroad and diagnosis of zygosity is rarely made. We did the analysis of clinical material on two sets of triplets. Diagnosis and analysis of zygosity were made in eleven ways about 57 items including blood type and RBC enzyme type, etc.
文摘The study of twins provides a unique opportunity to evaluate the genetic and nongenetic contributions to a variance of multitude of traits.Twin research has revealed the impact of a strong genetic component to the variance of many dental traits.The present study was conducted to assess the dental caries and oral hygiene status in monozygotic(MZ)and dizygotic(DZ)twins.A random sample of 129 MZ and DZ twin pairs aged 6-15 years were included in this study.The zygosity of twin pairs was initially recorded by facial appearance and later determined by dermatoglyphics.Dental caries and oral hygiene status were recorded in these twin pairs and the data were subjected to statistical analysis.No significant difference was observed in the mean decayed,extracted due to caries and filled teeth(deft)in primary dentition and decayed,missing and filled(DMFT)in permanent dentition scores and oral hygiene status in MZ and DZ twin pair&The similarity observed in oral health status reflects the effects of a shared environment in MZ and DZ twin pairs.
