Vectorette PCR isolation of microsatellite repeat sequences using anchored dinucleotide repeat primers.

نویسندگان

  • N J Lench
  • A Norris
  • A Bailey
  • A Booth
  • A F Markham
چکیده

We have developed a vectorette PCR approach to provide an improved method for isolation of microsatellite repeats. The modified procedure relies on PCR amplification using a vectorette-specific primer in combination with one of a panel of anchored dinucleotide repeat primers. The target DNA to be screened for microsatellite sequences can be from YAC, P1, cosmid, bacteriophage or plasmid clones. We have used this technique to isolate novel, polymorphic microsatellite repeats from clones containing the amelogenin gene (AMGX) located on human chromosome Xp22.3.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Designing Of Degenerate Primers-Based Polymerase Chain Reaction (PCR) For Amplification Of WD40 Repeat-Containing Proteins Using Local Allignment Search Method

Degenerate primers-based polymerase chain reaction (PCR) are commonly used for isolation of unidentified gene sequences in related organisms. For designing the degenerate primers, we propose the use of local alignment search method for searching the conserved regions long enough to design an acceptable primer pair. To test this method, a WD40 repeat-containing domain protein from Beauveria bass...

متن کامل

Microsatellite Isolation and Characterization in Pomegranate (Punica granatum L.)

Development of microsatellite markers has been an increasing trend in crop genetic studies because oftheir applicability in breeding programs. Here we report the development of inter simple sequencerepeat (SSRs) in pomegranate (Punica granatum L.) using an enrichment method that makes use of magneticbeads. Enriched genomic libraries with AG and ATG microsatellite motifs were c...

متن کامل

MS-IRS PCR: a simple method for the isolation of microsatellites.

Microsatellites (MS) are excellent genetic markers because of their prevalence in eukaryotic genomes, ease of typing by PCR and high degree of polymorphism. MS-based genetic maps of the human and mouse genomes have been constructed with average intermarker interval sizes of 1.6 cM and 0.42 cM, respectively (1,6). While these maps are essential resources for gene mapping, their resolution may no...

متن کامل

Improved fidelity of thermostable ligases for detection of microsatellite repeat sequences using nucleoside analogs.

Microsatellite repeats consisting of dinucleotide sequences are ubiquitous in the human genome and have proven useful for linkage analysis, positional cloning and forensic identification purposes. In this study, the potential of utilizing the ligase detection reaction for the analysis of such microsatellite repeat sequences was investigated. Initially, the fidelity of thermostable DNA ligases w...

متن کامل

Rapid development of gene-tagged microsatellite markers from bacterial artificial chromosome clones using anchored TAA repeat primers.

We developed a technique to improve the efficiency of producing TAA repeat microsatellite markers linked to interspecific conserved genes. Template DNA was prepared from cultures derived from single bacterial artificial chromosome (BAC) colonies using a simple alkaline lysis miniprep. The presence of conserved genes in each BAC clone was verified by sequencing with gene-specific primers. The BA...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Nucleic acids research

دوره 24 11  شماره 

صفحات  -

تاریخ انتشار 1996