Development of a Quick SSCP Method to Detect the Common O Variant Alleles
Location
CSU Ballroom
Start Date
21-4-2008 1:00 PM
End Date
21-4-2008 3:00 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
In conventional blood analysis, there is only one O blood group type. However, on the genetic level, multiple types of O alleles exist with different DNA sequences. Some of the uncommon alleles which contain single nucleotide polymorphisms (SNPs) have caused complications in blood transfusion and organ transplants. ABO genotyping has also been useful in anthropological studies since some alleles are more prevalent in certain cultures. The purpose of this project was to investigate the feasibility of using SSCP (single stranded conformational polymorphisms) to identify ABO genotypes, including those containing the rarer 0303 and 0209 alleles. Allele specific primers (ASP) were used to confirm the presence of the Al0l, Bl0l, 0101, 0201, 0209, and 0303 alleles. In the initial study, ten blood samples were analyzed using the ASP and SSCP methods. The DNAs were isolated using the Gentra Generation Capture Column Kit. The ASP polymerase chain reaction (PCR) procedure was designed to detect the G802A and the A2 97G SNPs, and the SSCP PCR method amplified cDNA segments from these regions. All PCR products were run on 15% polyacrylamide gels, but the SSCP DNAs were first denatured in formamide. Both gels were silver stained and analyzed. One of 10 samples was positive for the 0303 allele and this showed a unique pattern on the SSCP gel. Four different SSCP patterns were detected for this region and six different genotype patterns were found for the 297 region. Further SSCP method development could provide a much faster and cheaper alternative to ABO genotyping.
Development of a Quick SSCP Method to Detect the Common O Variant Alleles
CSU Ballroom
In conventional blood analysis, there is only one O blood group type. However, on the genetic level, multiple types of O alleles exist with different DNA sequences. Some of the uncommon alleles which contain single nucleotide polymorphisms (SNPs) have caused complications in blood transfusion and organ transplants. ABO genotyping has also been useful in anthropological studies since some alleles are more prevalent in certain cultures. The purpose of this project was to investigate the feasibility of using SSCP (single stranded conformational polymorphisms) to identify ABO genotypes, including those containing the rarer 0303 and 0209 alleles. Allele specific primers (ASP) were used to confirm the presence of the Al0l, Bl0l, 0101, 0201, 0209, and 0303 alleles. In the initial study, ten blood samples were analyzed using the ASP and SSCP methods. The DNAs were isolated using the Gentra Generation Capture Column Kit. The ASP polymerase chain reaction (PCR) procedure was designed to detect the G802A and the A2 97G SNPs, and the SSCP PCR method amplified cDNA segments from these regions. All PCR products were run on 15% polyacrylamide gels, but the SSCP DNAs were first denatured in formamide. Both gels were silver stained and analyzed. One of 10 samples was positive for the 0303 allele and this showed a unique pattern on the SSCP gel. Four different SSCP patterns were detected for this region and six different genotype patterns were found for the 297 region. Further SSCP method development could provide a much faster and cheaper alternative to ABO genotyping.
Recommended Citation
Karanjeet, Kul and Casey Malone. "Development of a Quick SSCP Method to Detect the Common O Variant Alleles." Undergraduate Research Symposium, Mankato, MN, April 21, 2008.
https://cornerstone.lib.mnsu.edu/urs/2008/poster-session-B/28
