A simple PCR method for screening cDNA libraries.
نویسندگان
چکیده
منابع مشابه
PCR-based Method for Screening Libraries
One of the fundamental techniques of molecular biology is the isolation of a rare clone from a complex mixture of DNA sequences contained within a library. The isolation of cDNA and genomic clones from highly complex libraries is often an early step in studies aimed at understanding basic biological processes as well as in applied biological research. In a typical genomic library with an averag...
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Screening a cDNA library is a common practice for cloning genes of interest in many laboratories. Success of screening is largely dependent on the abundance of cDNAs representing genes of interest and the availability of suitable probes. The procedure involves several rounds of screening to obtain true-positive clones, which might take a week or longer. Typically, each round of screening requir...
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The common procedure for isolating genes of interest is to screen cDNA or genomic libraries with nucleic acid probes by filter hybridization (1). However, this is a time consuming and laborintensive process, especially when the complexity of the library is great and there is a large number of bactenophage plaques to screen. Here we describe the use of the polymerase chain reaction (PCR) (2) to ...
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The systematic sequencing of cDNA libraries is an efficient approach for the identification of new genes, but the presence of abundant mRNAs is often a major problem. This paper describes a very simple method of "semi-multiplex PCR" that allows specific identification of such abundant transcripts before DNA sequencing without using nonrepresentative subtracted libraries. The PCR utilizes a seri...
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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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ژورنال
عنوان ژورنال: Genome Research
سال: 1994
ISSN: 1088-9051
DOI: 10.1101/gr.4.1.46