Development and Validation of the AmpFlSTRR Yfilertm PCR Amplification Kit: A Male Specific, Single Amplification 17 Y-STR Multiplex System*

2006 ◽  
Vol 51 (1) ◽  
pp. 64-75 ◽  
Author(s):  
Julio J. Mulero ◽  
Chien Wei Chang ◽  
Lisa M. Calandro ◽  
Robert L. Green ◽  
Yixin Li ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Male Specific ◽  
Development And Validation ◽  
Multiplex System

scholarly journals New Male Specific Markers for Hop and Application in Breeding Program

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Andreja Čerenak ◽  
Zala Kolenc ◽  
Petra Sehur ◽  
Simon P. Whittock ◽  
Anthony Koutoulis ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Dna Sequences ◽  
Low Cost ◽  
Pcr Amplification ◽  
Breeding Program ◽  
Phenotypic Data ◽  
World Collection ◽  
Crude Sample ◽  
Male Sex ◽  
Male Specific

Abstract Male specific DNA sequences were selected from a Diversity Arrays Technology (DArT) mapping study to evaluate their suitability for determination of the sex phenotype among young seedlings in a hop (Humulus lupulus L.) breeding program. Ten male specific DArT markers showed complete linkage with male sex phenotype in three crossing families. Following optimization, four were successfully converted into PCR markers and a multiplex PCR approach for their use was developed. Among 197 plants (97 from the world collection; 100 from three segregating families), 94–100% positive correlation with sex phenotypic data was achieved for the single PCR amplification, whereas the multiplex approach showed 100% correlation. To develop a fast and low-cost method, crude sample multiplex PCR was evaluated in 253 progenies from 14 segregating populations without losing accuracy. The study describes, for the first time, the routine application of molecular markers linked to male sex in an intensive Slovenian hop breeding program. The methods described could be employed for screening of sex at the seedling stage in other hop programs worldwide, thereby saving resources for desirable female plants.

369 AMPLIFICATION OF THE SRY GENE FOR SEXING OF BUFFALO (BUBALUS BUBALIS) EMBRYOS USING NESTED MULTIPLEX PCR

2007 ◽  
Vol 19 (1) ◽  
pp. 300
Author(s):  
M. Zhang ◽  
Q. Fu ◽  
W. S. Qin ◽  
H. Y. Zheng ◽  
Y. Q. Lu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Internal Control ◽  
Pcr Amplification ◽  
Single Copy ◽  
Bubalus Bubalis ◽  
Sex Identification ◽  
Sry Gene ◽  
Embryo Sex ◽  
Pcr Method ◽  
Male Specific

In mammals, the Y chromosome-linked SRY gene is responsible for male sex determination. Therefore, a logical approach for embryo sex identification is to amplify the male-specific single-copy SRY gene. The objective of this study was to design specific primers for amplification of buffalo SRY gene and develop a reliable PCR method for sex identification of buffalo embryos. Genomic DNA was extracted from swamp buffalo (Bubalus bubalis) peripheral blood. A pair of primers based on the sequence of Holstein bovine SRY gene (forward, 52-GTTTGCCTTATGGATTTATT-32; reverse, 52-TCTACTTTAGCCTATTTG-32) was used to amplify whole buffalo SRY gene. This amplified fragment was isolated and constructed into plasmids for sequencing. Two pairs of primers, S1/S2 (forward, 52-CCATGAACGCCTTCATTTTGTG-32; reverse, 52-ACGAGGTCGATATTTATAGC CC-32) and S3/S4 (forward, 52-AAGCAGCTGGGCTATGAGTGGAA-32; reverse, 52-ACGAGGTCGATATTTATAGCCC-32), were designed based on the SRY sequence above. Simultaneously, the G3PDH gene was amplified to serve as an internal control for both male and female embryos. A multiplex-nested-PCR system was optimized by varying the following parameters individually: concentration of Mg2+, dNTPs, primers, and different cycles number. Twenty-seven IVF morulae were identified with the optimal PCR procedure after biopsy. Accuracy of PCR amplification was verified by dot blotting. The sex of 24 embryos fertilized with Y-sperm separated by flow cytometry was also examined. Results indicated that the optimal procedure of Nested-Multiplex-PCR consisted of 1.5 mM Mg2+, 100 �M dNTPs, 0.5 �M SA3/SA4 primers, and 0.25 �M GA3/GA4 primers, and 35 cycles. Accuracy of identification was 100% for 27 IVF morulae; 14 were judged as males and 13 were females. The result of blotting confirmed that the accuracy of amplification was 100%. The proportion of males was 83.3% (20/24) in embryos fertilized with Y-sperm. This confirms that the PCR system targeting the SRY gene can be used for accurate sex identification of buffalo embryos. This study was supported by grants from the Foundation of Guangxi Key Laboratory for Subtropical Bio-Resource Conservation and Utilization (SB0403) and the Guangxi Department of Science and Technology (0626001-3-1).

scholarly journals Identification and Validation of a New Male Sex-Specific ISSR Marker in Pointed Gourd (Trichosanthes dioicaRoxb.)

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Sinchan Adhikari ◽  
Soumen Saha ◽  
Tapas Kumar Bandyopadhyay ◽  
Parthadeb Ghosh
Keyword(s):  
Pcr Amplification ◽  
Specific Marker ◽  
Reliable Technique ◽  
Sequence Characterization ◽  
Gender Determination ◽  
Pooled Dna ◽  
Sex Marker ◽  
Pointed Gourd ◽  
Male Specific ◽  
Issr Primers

The aim of the present study was to develop a genetic sex marker for the pointed gourd (Trichosanthes dioicaRoxb.) to allow gender determination at any stage in the life cycle. Screening of genomic DNA with intersimple sequence repeat (ISSR) primers was used to discover sex-specific touch-down polymerase chain reaction (Td-PCR) amplification products. Using pooled DNA from male and female genotypes and 42 ISSR primers, a putative male specific marker (~550 bp) was identified. DNA marker specific to male is an indication of existence of nonepigenetic factors involved in gender development in pointed gourd. The ISSR technique has proved to be a reliable technique in gender determination of pointed gourd genotypes at the seedling phenophase. The sex marker developed here could also be used as a starting material towards sequence characterization of sex linked genes for better understanding the developmental as well as evolutionary pathways in sexual dimorphism.

scholarly journals Development and validation of a male specific body checking questionnaire

2009 ◽  
pp. NA-NA ◽  
Author(s):  
Tom Hildebrandt ◽  
D. Catherine Walker ◽  
Lauren Alfano ◽  
Sherrie Delinsky ◽  
Katie Bannon
Keyword(s):  
Body Checking ◽  
Male Specific ◽  
Development And Validation

Development and validation of I-DNA1: a 15-Loci multiplex system for identity testing

2011 ◽  
Vol 125 (5) ◽  
pp. 685-694 ◽  
Author(s):  
A. Odriozola ◽  
J. M. Aznar ◽  
D. Celorrio ◽  
M. L. Bravo ◽  
J. J. Builes ◽  
...  
Keyword(s):  
Identity Testing ◽  
Development And Validation ◽  
Multiplex System

scholarly journals A multiplex-system to target 16 male-specific and 15 autosomal genetic markers for orang-utans (genus: Pongo)

2010 ◽  
Vol 2 (S1) ◽  
pp. 153-158 ◽  
Author(s):  
Pirmin Nietlisbach ◽  
Alexander Nater ◽  
Maja P. Greminger ◽  
Natasha Arora ◽  
Michael Krützen
Keyword(s):  
Genetic Markers ◽  
Male Specific ◽  
Multiplex System

Development and validation for identity testing of I-DNADuo, a combination of I-DNA1 and a new multiplex system, I-DNA2

2011 ◽  
Vol 126 (1) ◽  
pp. 167-172 ◽  
Author(s):  
Adrian Odriozola ◽  
Jose María Aznar ◽  
David Celorrio ◽  
Maria Luisa Bravo ◽  
Juan Jose Builes ◽  
...  
Keyword(s):  
Identity Testing ◽  
Development And Validation ◽  
Multiplex System

scholarly journals Development and Validation of a Forensic Multiplex System With 38 X-InDel Loci

2021 ◽  
Vol 12 ◽  
Author(s):  
Ling Chen ◽  
Xiyong Pan ◽  
Yuan Wang ◽  
Weian Du ◽  
Weibin Wu ◽  
...  
Keyword(s):  
Hemoglobin Concentration ◽  
Peak Height ◽  
Parameter Analysis ◽  
Single Polymerase Chain Reaction ◽  
Good Tolerance ◽  
Discrimination Capability ◽  
Polymerase Chain ◽  
Template Dna ◽  
Development And Validation ◽  
Multiplex System

In the present study, a novel multiplex system, AGCU X-InDel 38 kit, was designed to amplify 38 X-InDel markers and amelogenin in a single Polymerase Chain Reaction (PCR). To demonstrate the suitability and efficiency for forensic applications, a series of validation experiments were conducted, including sensitivity, species specificity, reproducibility, stability, case samples, balance of peak height, size precision, as well as allele frequency and forensic parameter analysis. The results showed that AGCU X-InDel 38 kit was capable to get full profiles even with 62.5 pg of template DNA, and full profiles can be obtained when hematin concentration ≤25 μmol/L, or hemoglobin concentration ≤50 μmol/L, showing good tolerance to six common inhibitors. Moreover, the analyzed case samples indicated that AGCU X-InDel 38 kit had better performance for degraded and trace DNA samples. The 200 unrelated males from Guangdong Han population showed that the combined PDMale and PDFemale were both more than 0.999999999, and the combined MECKrüger, MECKishida, and MECDesmarais  Duo were 0.999369481, 0.999999917, and 0.999941556, respectively. Robust discrimination capability of this novel multiplex system could be demonstrated through the high values of forensic parameters. In conclusion, AGCU X-InDel 38 kit is sensitive, precise, reproducible, and highly informative and could be used as a complementary tool for complex and challenging kinship cases.

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