Primer selection guide for use with GoScript® Reverse Transcription Mixes29.08.2018
To initiate reverse transcription, reverse transcriptases require a short DNA oligonucleotide called a primer to bind the RNA template and serve as a starting point for synthesis of a new strand. Depending on the RNA template and the downstream applications, primers of three basic types are available: oligo(dT) primers, random primers and gene-specific primers. Each primer binds to a complementary sequence on the RNA template to facilitate reverse transcription, the difference between these options is the sequence that they bind. Gene specific primers bind target sequences contained within a single mRNA of interest and only that region is amplified; these primers are often used for one-step RT-qPCR reactions. Oligo(dT) primers amplify only mRNAs containing a poly(A) tail, since that is where the primer binds to promote reverse transcription. Random primers amplify most RNA species, including degraded RNA and viral genomes. In some cases both oligo(dT) and random primers can be used to combine the benefits of both primer types. We compared the GoScript™ Reverse Transcription Mixes, Oligo(dT) and Random Primers, to a 50/50 mixture of the reagents for ability to reverse transcribe three RNA transcripts.
Hook, B. and Lewis, S. Primer selection guide for use with GoScript® Reverse Transcription Mixes. [Internet] June 2017; tpub_185. [cited: year, month, date]. Available from: http://worldwide.promega.com/resources/pubhub/primer-selection-guide-for-use-with-goscript-reverse-transcription-mixes/