Herein, we developed a simple, label-free, isothermal, and ultrasensitive colorimetric method for amplified detection of DNA on the basis of a new quadratic amplification strategy.
In this work, by taking the uniqueness of Exonuclease III and DNA metallization, we demonstrate a facile turn-on fluorescent method for amplified detection of biothiols at picomolar concentration.
In this new method, electrophoretic analysis of amplified products is replaced with photometric analysis of a probed oligonucleotide array.
The improved yield over the BDSmart method is notable considering that both of these methods involve production of amplified sense RNA (Table 2).
Batch mean-centering (see methods) of the amplified and unamplified datasets was found to dramatically increase the correspondence comparison across the datasets, with 100% of probesets having fold changes within two-fold between experimental branches (Fig. 1B black dots, Table 1).
Extraction with guanidinium-phenol and Lithium Chloride precipitation are the optimal methods for purification of amplified RNA and labelled aRNA respectively.
A combined method of sequence characterized amplified regions (SCAR) primers and Touchdown-PCR was used for the development of a female DNA marker in Pistacia vera L. The random decamer primer OPO-08 amplified a 905-bp fragment in all female trees, but also in several males.
Recently, the high-resolution method of specific length amplified fragment sequencing (SLAF-seq), which is based on RRLs and high-throughput sequencing for large-scale genotyping and de novo SNP discovery, was reported [ 28].
These results not only agree with the current hypotheses regarding the mechanism of GBS/MFS but also concur with a recent study in which GBS/MFS strains and enteritis-control strains were screened for potential neuropathogenic markers using the high resolution comparative genomic method of high-throughput Amplified Fragment Length Polymorphism (htAFLP) [ 44].
In this setting, our method produces a population of amplified RNA having minimal and consistent amplification bias, orientation compatible with both spotted cDNA and oligonucleotide arrays, and quantity sufficient for several downstream assays from a single amplification.
After initial tests of five complexity reduction methods through gel electrophoresis of amplified products (data not presented), the ' PstI/ TaqI' method was chosen for library construction.
