Allele-Specific PGR and SSCP Analysis of ABO Variants
Location
CSU Ballroom
Start Date
28-4-2009 10:00 AM
End Date
28-4-2009 12:00 PM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Theresa Salerno
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
The ABO blood group system is the most important blood group system for various clinical applications. Some genetic variants of the four basic blood groups A, B, AB and typing that have been problematic in some cases of transfusion medicine. These variants were usually different by only one nucleotide and are called Single Nucleotide Polymorphisms (SNPs). The purpose of this study was to develop a quick and simple methodology called Allele Specific Polymerase Chain Reaction- Single Strand Conformation Polymorphism (ASPCR- SSCP) to discover novel variants of ABO alleles and especially new 0 variant alleles. This PCR-SSCP method amplified and separated DNA molecules based on subtle differences in single stranded DNA sequences and resulted in different banding patterns on a 15% gel following denaturation to single stranded forms. The ability to use allele specific amplification prior to SSCP analysis simplified patterns for heterozygotes. For this study DNA was isolated from blood samples using a column capture kit. Two new sets of primers were designed for both region 1 (exon 6) and region 3 (nucleotides 628- 940 of exon 7) of the ABO gene. DNAs were amplified by PCR technique and then analyzed on a 15% polyacrylamide gel following formamide denaturation. The variant represented by new banding pattern was fiirther identified using a Li-Cor DNA sequencer. This study showed that the PCR-SSCP method is an efficient and cost effective way for ABO genotyping to be used in applied research.
Allele-Specific PGR and SSCP Analysis of ABO Variants
CSU Ballroom
The ABO blood group system is the most important blood group system for various clinical applications. Some genetic variants of the four basic blood groups A, B, AB and typing that have been problematic in some cases of transfusion medicine. These variants were usually different by only one nucleotide and are called Single Nucleotide Polymorphisms (SNPs). The purpose of this study was to develop a quick and simple methodology called Allele Specific Polymerase Chain Reaction- Single Strand Conformation Polymorphism (ASPCR- SSCP) to discover novel variants of ABO alleles and especially new 0 variant alleles. This PCR-SSCP method amplified and separated DNA molecules based on subtle differences in single stranded DNA sequences and resulted in different banding patterns on a 15% gel following denaturation to single stranded forms. The ability to use allele specific amplification prior to SSCP analysis simplified patterns for heterozygotes. For this study DNA was isolated from blood samples using a column capture kit. Two new sets of primers were designed for both region 1 (exon 6) and region 3 (nucleotides 628- 940 of exon 7) of the ABO gene. DNAs were amplified by PCR technique and then analyzed on a 15% polyacrylamide gel following formamide denaturation. The variant represented by new banding pattern was fiirther identified using a Li-Cor DNA sequencer. This study showed that the PCR-SSCP method is an efficient and cost effective way for ABO genotyping to be used in applied research.
Recommended Citation
Gamage, Yohani K G V. "Allele-Specific PGR and SSCP Analysis of ABO Variants." Undergraduate Research Symposium, Mankato, MN, April 28, 2009.
https://cornerstone.lib.mnsu.edu/urs/2009/poster-session-C/20
