PCR Template Preparation for Capillary DNA Sequencing
نویسندگان
چکیده
منابع مشابه
PCR template preparation for capillary DNA sequencing.
Fluorescence-based capillary DNA sequencing has facilitated the early completion of several complex sequencing projects. While capillary systems offer great benefits in terms of ease of use and automation, we find that they are sufficiently different from slab gel separation methodologies, demanding re-examination of the protocols used to generate and use DNA sequencing templates. We have recen...
متن کاملPreparation of PCR products for DNA sequencing.
We demonstrate that routine PCR product analytical agarose gels can also serve as preparative gels for quick DNA template purification before sequencing. The band of interest is excised, placed into a Gel Nebulizer inside a Micropure separator and rapidly purified in a single centrifugation step. Gel-purified PCR product, suitable for manual and automated sequencing, is delivered within 10 min.
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Isolation of DNA for PCR is time-consuming and involves many reagents. The aim of this work was to optimise a rapid and easy PCR methodology without previous DNA isolation. Different strains of the phytopathogenic fungus Colletotrichum lindemuthianum were used. Protoplasts were generated using lytic enzymes under high incubation temperatures using different methodologies to obtain the template....
متن کاملSequencing of PCR-amplified DNA.
Alternatives for sequencing of PCR products essentially fall into one of two categories; generation of single-stranded DNA for sequencing or the direct sequencing of double-stranded product. Of the two alternatives, sequencing of double-stranded PCR products is likely to be of greatest immediate significance in terms of general applicability and rapidity. Double-stranded sequencing allows the u...
متن کاملPreparation of template DNA and labeling techniques.
A variety of probes can be used for in situ hybridization, depending on the application and the labeling strategy. In general, RNA probes (riboprobes) are now more commonly used. However, some laboratories may still prefer to use DNA probes for in situ hybridization, and a number of techniques for this type of labeling have been described in this chapter. The preparation of plasmid DNA and the ...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 2001
ISSN: 0736-6205,1940-9818
DOI: 10.2144/01303st05