A simple and extremely sensitive dual-amplification platform for the detection of DNA was developed combining signal amplification of conjugated polymers and recyclable autocatalytic hybridization of DNA.

Helicase-dependent amplification (HDA) is an isothermal in vitro DNA amplification method based upon the coordinated actions of helicases to separate double-stranded DNA and DNA polymerases to synthesize DNA. Previously, a mesophilic form of HDA (mHDA) utilizing the Escherichia coli UvrD helicase, DNA polymerase I Klenow fragment, two accessory proteins, MutL and single-stranded DNA-binding pro...

A clinical procedure for rapid detection of human immunodeficiency virus type 1 (HIV-1) by DNA amplification is demonstrated. The rapid procedure reduces handling requirements and amplification time and eliminates use of radioactivity for the detection of the amplification product. Total leukocyte lysates are the amplification substrates. Two conserved regions in the HIV-1 genome are amplified ...

The DNA amplification performed by terminal protein-primed replication systems has not yet been developed for its general use to produce high amounts of DNA linked to terminal protein (TP). Here we present a method to amplify in vitro heterologous DNAs using the Φ29 DNA replication machinery and producing DNA with TP covalently attached to the 5' end. The amplification requires four Φ29 protein...

The reported genome-walking methods still suffer from some deficiencies, such as cumbersome experimental steps, short target amplicon, or deep background. Here, a simple and practical fork PCR was proposed for genome-walking. employs primer set of three random oligomers to implement walking task. In primary PCR, the low-stringency amplification cycle mediates binding places on genome, producing...

Single cell whole genome amplification is susceptible to amplification biases that impact the accuracy of single cell sequencing data. To address this, we have developed a microfluidic device for the isolation and purification of genomic DNA from individual cells. The device uses a micropillar array to physically capture single cells and its chromosomal DNA upon extraction. The extracted DNA is...

We describe conditions for rolling-circle amplification (RCA) of individual DNA molecules 5–7 kb in size by >109-fold, using 29 DNA polymerase. The principal difficulty with amplification of small amounts of template by RCA using 29 DNA polymerase is ‘‘background’’ DNA synthesis that usually occurs when template is omitted, or at low template concentrations. Reducing the reaction volume while k...

Introduction • The diagnostic use of indicator-dilution technics in the study of congenital heart disease dates, for practical purposes, from 1950 with the nearly simultaneous description of methods for continuous recording of the concentration of Evans blue dye from Johns Hopkins University and from the Mayo Clinic." The diagnostic applications of indicatoi'-dilution technics have been intensi...

methylated and un-methylated control dna is an important part of dna methylation studies. although human and mouse dna methylation control sets are commercially available, in case of methylation studies on other species such as animals, plants, and bacteria, control sets need to be prepared. in this paper a simple method of generating methylated and un-methylated control dna is described. whole...