Abstract
BackgroundThe ability to transport and store DNA at room temperature in low volumes has the advantage of optimising cost, time and storage space. Blood spots on adapted filter papers are popular for this, with FTA (Flinders Technology Associates) Whatman™TM technology being one of the most recent. Plant material, plasmids, viral particles, bacteria and animal blood have been stored and transported successfully using this technology, however the method of porcine DNA extraction from FTA Whatman™TM cards is a relatively new approach, allowing nucleic acids to be ready for downstream applications such as PCR, whole genome amplification, sequencing and subsequent application to single nucleotide polymorphism microarrays has hitherto been under-explored.FindingsDNA was extracted from FTA Whatman™TM cards (following adaptations of the manufacturer’s instructions), whole genome amplified and subsequently analysed to validate the integrity of the DNA for downstream SNP analysis. DNA was successfully extracted from 288/288 samples and amplified by WGA. Allele dropout post WGA, was observed in less than 2% of samples and there was no clear evidence of amplification bias nor contamination. Acceptable call rates on porcine SNP chips were also achieved using DNA extracted and amplified in this way.ConclusionsDNA extracted from FTA Whatman cards is of a high enough quality and quantity following whole genomic amplification to perform meaningful SNP chip studies.
Highlights
The ability to transport and store Deoxyribonucleic acid (DNA) at room temperature in low volumes has the advantage of optimising cost, time and storage space
The optimum for which the punches were heated to was deemed to be 15 seconds and the optimum length of time that the punches were washed for was deemed to be 5 seconds
This study has successfully demonstrated the efficacy of Flinders Technology Associates (FTA) WhatmanTM cards as a tool for storing porcine genomic DNA at room temperature largely supporting the manufacturer’s claims
Summary
The ability to transport and store DNA at room temperature in low volumes has the advantage of optimising cost, time and storage space. Plasmids, viral particles, bacteria and animal blood have been stored and transported successfully using this technology, the method of porcine DNA extraction from FTA WhatmanTM cards is a relatively new approach, allowing nucleic acids to be ready for downstream applications such as PCR, whole genome amplification, sequencing and subsequent application to single nucleotide polymorphism microarrays has hitherto been under-explored. Findings: DNA was extracted from FTA WhatmanTM cards (following adaptations of the manufacturer’s instructions), whole genome amplified and subsequently analysed to validate the integrity of the DNA for downstream SNP analysis. DNA was successfully extracted from 288/288 samples and amplified by WGA. Acceptable call rates on porcine SNP chips were achieved using DNA extracted and amplified in this way
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
