Abstract:
:Differential display of mRNA is a simple, sensitive and powerful method to identify differentially expressed gene fragments. The main drawback of differential display is the lack of reproducibility and the inability to read and compare complex gels. This issue results from employing unoptimized primer combinations and non-specific amplification, most likely due to unavoidable low annealing temperatures. In order to display most of the expressed transcripts (80-120 bands/lane), 26 different 5' primers were used in conjunction with nine different 3' poly (dT) primers. These primer combinations, used with the optimized annealing temperature for each set of primers, produced highly reproducible bands. BSA has a direct effect on the number of bands resolved. Variations in ramping time (9-40 s) had little or no effect on the resolution and reproducibility of differential display.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Malhotra K,Foltz L,Mahoney WC,Schueler PAdoi
10.1093/nar/26.3.854subject
Has Abstractpub_date
1998-02-01 00:00:00pages
854-6issue
3eissn
0305-1048issn
1362-4962pii
gkb173journal_volume
26pub_type
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