Minimizing false positives in differential display.
نویسندگان
چکیده
منابع مشابه
Reverse slot blot for the verification of cDNAs identified through differential display.
Differential display, first described by Liang and Pardee (4), has become a powerful screening tool for the identification of cellor stage-specific gene expression. The technique relies on efficient reverse transcription of mRNA and subsequent amplification of this cDNA with as many random primer combinations as possible. RT-PCR products are separated on denaturing sequencing gels, visualized t...
متن کاملElimination of false positives generated through PCR re-amplification of differential display cDNA.
Differential display (DD) is a powerful molecular tool that allows the identification and subsequent isolation of transcripts differentially expressed between biological samples, for example, between undifferentiated and differentiated cells, between different tissues or in one tissue at different stages of development. However, significantly high rates of apparent false positives have been rep...
متن کاملModified differential display technique that eliminates radioactivity and decreases screening time.
Several techniques are available that detect variations in gene expression between cellular populations. These include subtractive hybridization (SH), differential colony hybridization (DCH) and mRNA differential display, all based on the analysis of mRNA. The first two techniques, however, are limited because they require large amounts of mRNA for SH or several rounds of screening for DCH. Dif...
متن کاملIdentification and verification of differential display cDNAs using gene-specific primers and hybridization arrays.
An accurate and streamlined approach to differential display (DD) band identification and verification is described. To minimize false positives, the strategy avoids the use of impure Northern blot probes obtained from PCR-amplified DD bands. To increase throughput, the cloning of DD bands is replaced by a gene-specific primer approach, and hybridization arrays are used in place of Northern blo...
متن کاملImproved Procedure for Screening Expression Libraries for Novel Autoantigens
The standard method for immunoscreening of a cDNA expression library is time-consuming becauseof the production of a large proportion of false positives during the first and second round of screening.This problem is more important when a sensitive chemiluminescence detection system is used. Due tothe high sensitivity of the detection system, there is a need to avoid false posi...
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عنوان ژورنال:
- BioTechniques
دوره 24 5 شماره
صفحات -
تاریخ انتشار 1998