A rapid protocol for the authentication of isolated differential display RT-PCR CDNAs. |
| |
Authors: | G Miele R Slee J Manson M Clinton |
| |
Institution: | Division of Development & Reproduction, Roslin Institute, Midlothian, Scotland, UK. |
| |
Abstract: | The most time-consuming and problematic step in the overall DDRT-PCR technique is the confirmation that the isolated cDNA clone represents a differentially expressed gene. We have previously suggested that the majority of apparent false positives generated by DDRT-PCR do in fact result from the PCR reamplification of cDNA species which co-migrate with the cDNA of interest, and we have outlined a procedure to effectively eliminate these from further study. However, in situations where RNA is limiting, it is still desirable to confirm that a purified cDNA amplicon does, in fact, represent the originally observed differentially expressed gene prior to embarking on expression studies. |
| |
Keywords: | |
|
|