PCR-Based Method for Isolation of Full-Length Clones and Splice Variants from cDNA Libraries
نویسندگان
چکیده
منابع مشابه
PCR-based method for isolation of full-length clones and splice variants from cDNA libraries.
Most cDNA library screening procedures do not distinguish between full-length and incomplete clones and therefore may yield incomplete cDNA fragments. Thus, there is a widespread need for a method allowing the efficient selection of full-length clones. I present a rapid, PCR-based method that allows the simultaneous screening of > 10(6) cDNAs. The longest cDNA is identified in the first step so...
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Pseudogenes, alternative transcripts, noncoding RNA, and polymorphisms each add extensive complexity to the mammalian transcriptome and confound estimation of the total number of genes. Despite advanced algorithms for gene prediction and several large-scale efforts to obtain cDNA clones for all human open reading frames (ORFs), no single collection is complete. To enhance this effort, we have d...
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Obtaining full-length or even near full-length cDNA clones has been a time-consuming and labor-intensive step in the analysis of cloned genes. Recent methods which use PCR as a preparative tool for cloning (1,2) have made this step considerably more rapid, but may introduce sequence errors in the resulting clones due to numerous sequential rounds of in vitro replication. We describe a method fo...
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BACKGROUND cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) is a commonly used technique for genome-wide expression analysis that does not require prior sequence knowledge. Typically, quantitative expression data and sequence information are obtained for a large number of differentially expressed gene tags. However, most of the gene tags do not correspond to full-length (FL) coding seque...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1997
ISSN: 0736-6205,1940-9818
DOI: 10.2144/97223st02