منابع مشابه
DNA sequencing using Taq polymerase.
Three DNA polymerases, namely E.coli DNA polymerase 1 (Klenow), reverse transcriptase and T7 DNA polymerase (sequenase), are commonly used for DNA sequencing by the chain termination method of Sanger and colleagues [1). However, the secondary structure of the DNA template can impede the progress of all three polymerases. I have developed a novel procedure in which the thermostable polymerase of...
متن کاملHeterozygote and mutation detection by direct automated fluorescent DNA sequencing using a mutant Taq DNA polymerase.
We describe a method for direct cycle sequencing of PCR fragments amplified from genomic DNA or cDNA. DNA sequencing template is amplified using PCR and oligonucleotide primers flanking the region of interest. The amplified fragment is directly cycle sequenced using fluorescent sequencing primers, Sanger dideoxy sequencing chemistry and an enzyme mixture of a mutant Taq DNA polymerase and therm...
متن کاملCycle Sequencing of Filamentous Phage DNA Using a Biotinylated Primer and ∆Taq DNA Polymerase
We describe a rapid nonradioactive, double-stranded phage DNA sequencing method using ∆Taq DNA polymerase in cycle reaction for phage peptide-display library screening. This procedure is specific, rapid, sensitive and safe for the sequencing of large numbers of phage peptide-display colonies. In addition, thermal cycle sequencing with this chemiluminescent image detection protocol provides an i...
متن کاملSimplified template preparation and improved direct sequencing using Taq polymerase.
A streamlined version of direct dideoxy sequencing is presented that includes template preparation as well as sequencing protocols. The method is used routinely to sequence double-stranded PCR products after minimal purification with one of the primers used in amplification. Either 35S or 32P labeling can be used with equally good results.
متن کاملDNA sequencing using polymerase substrate-binding kinetics
Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing. However, whole-genome sequencing is still costly and complex for diagnostics purposes. In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest. Routine clinical use of targeted NGS mandates inexpensive instruments, fast turna...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1988
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/16.22.10915