High and low annealing temperatures increase both specificity and yield in touchdown and stepdown PCR.

Single-step PCR optimization using touchdown and stepdown PCR programming.

Improved repetitive-element PCR fingerprinting of Salmonella enterica with the use of extremely elevated annealing temperatures.

Modified thermal cycling conditions were explored in an effort to improve the reproducibility and resolving power of repetitive-element PCR (rep-PCR) fingerprinting. Assay performance was rigorously evaluated under standard and modified cycling conditions, using as a test set 12 strains putatively representing 12 serovars of Salmonella enterica. For all three fingerprint types (ERIC2, BOXA1R, a...

Recrystallization Behavior of Deep Drawing Low Carbon Steel Sheets Produced by Mobarakeh Steel Plant

In the present work the recrystallization behavior of deep drawing grade Al-killed low carbon steel sheets produced by Mobarakeh Steel Plant (MSP) has been studied. Salt bath isothermal annealing has b...

A Nested-Splicing by Overlap Extension PCR Improves Specificity of this Standard Method

Background: Splicing by overlap extension (SOE) PCR is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function. Objectives: We introduced a nested-SOE-PCR (N –SOE-PCR) in order to increase the specificity and generating mutations in a gene by SOE-PCR.   Materials and Methods: Genomic DNA from Bacillus thermo...

Maximizing sensitivity and specificity of PCR by pre-amplification heating

We have found that assembling the reaction mixture at a temperature greater than the annealing temperature improved both product yield and specificity of PCR. When reactions were maintained at 70°C in a dry heating block during addition of denatured samples to aliquotted reagent master mix, a reproducible increase in product yield was observed compared to duplicates maintained at room temperatu...