A simple PCR procedure for discovering microsatellites from small insert libraries
نویسندگان
چکیده
Microsatellite discovery from genomic libraries is tedious because of the low number of clones that contain inserts and costly because of screening methodologies. A new procedure for screening clones for microsatellite DNA is described herein. Instead of colony hybridization, a polymerase chain reaction (PCR) with two vector standard primers and one synthesized repeat primer was used to directly screen colonies. PCR of colonies that produced a strong smear in gels contained the desired motif, whereas a single strong band indicated the lack of the desired motif. This simple screening method is a cost-effective way to identify microsatellite-containing colonies.
منابع مشابه
Rapid PCR-based characterization of sequences flanking microsatellites in large-insert libraries.
Microsatellite polymorphisms are widely being employed as a major tool in genome analysis. Analysis of length polymorphisms in the PCR depends on the elucidation of the sequence of their flanking regions. Different methods have been developed for the efficient characterization of the flanking sequences. Small insert genomic libraries directly suitable for sequencing of the entire insert and hig...
متن کاملIncreased yield of tri- and tetranucleotide heterospecific microsatellites from unenriched small-insert libraries.
متن کامل
Flow sorting of mitotic chromosomes in common wheat (Triticum aestivum L.).
The aim of this study was to develop an improved procedure for preparation of chromosome suspensions, and to evaluate the potential of flow cytometry for chromosome sorting in wheat. Suspensions of intact chromosomes were prepared by mechanical homogenization of synchronized root tips after mild fixation with formaldehyde. Histograms of relative fluorescence intensity (flow karyotypes) obtained...
متن کاملانتخاب اختصاصی فاژهای حاوی قطعه آنتیبادی به کمک پروتئین A در تکنیک کتابخانه آنتیبادی فاژ
Background and purpose: Antibody phage display library is a powerful in vitro technology for production of recombinant antibody fragments against a wide variety of antigens. However, the presence of insert-free clones in the phage libraries limited the specific enrichment of antibody fragments in many studies. The aim of this study was to protein A-aided recovery of insert-containing phages in ...
متن کاملA Set of 20 New SSR Markers Developed and Evaluated in Mandevilla Lindl.
Mandevilla is an ornamental crop with a bright future worldwide because of its high commercial acceptance and added value. However, as with most ornamental species, there are few molecular tools to support cultivar breeding and innovation. In this work, we report the development and analysis of 20 new Simple Sequence Repeat (SSR) markers in Mandevilla. Microsatellites were isolated from two enr...
متن کامل