T7 Amplification for Generating Single Stranded Target for Hybridization to Allele-Specific DNA Microarrays
Poster Feb 01, 2007
Jesper Petersen, Lena Poulsen, Henrik Birgens and Martin Dufva
Amplified RNA (aRNA) synthesized by T7 linear amplification is frequently used for gene expression analysis on microarrays. Single stranded DNA (ssDNA) is however mostly used as target in microarray based genotyping assays, although there are several advantages with aRNA. Here, we compared the performance of aRNA and ssDNA as targets in a genotyping assay, comprising a DNA microarray of allele specific oligo probes specific for thalassemic mutations.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE