PCR-based screening of YAC clones without DNA extraction

DNA extraction method for screening yeast clones by PCR.

A simple procedure for isolating yeast DNA suitable for use as a template for PCR amplification is described. SDS treatment alone is sufficient for extraction of chromosomal DNA from yeast cells. Cells of a yeast colony are suspended in a small volume (about 20 microL) of a 0.25% SDS solution, mixed vigorously and centrifuged. The supernatant can be directly used as a template after dilution to...

YAC Library Pooling Scheme for PCR-Based Screening

The PCR is a rapid method for screening a library of clones for the presence of clones containing an STS, Usually the library is divided into pools of clones, and the PCR is run on each pool. The problem we address here is to design efficient and robust pooling schemes for such PCR-based screening. Two questions are relevant: (1) Given an arbitrary unique sequence, how should one pool a library...

Digital PCR for direct quantification of viruses without DNA extraction

DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous m...

Diagnosis of mucocutaneous herpetic infections by PCR without DNA extraction.

Direct PCR from whole blood, without DNA extraction.

Typically DNA used in PCR assays is usually extracted according to die phenol-chloroform method (1) or by an alternative 'saltingout' rapid purification (2). Moreover partially purified DNA obtained with rapid procedures have been reported to be suitable for PCR amplification (3). We describe here a more simple and efficient method to amplify DNA directly from whole blood samples without any pu...