Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing

Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing.

Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subseque...

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*

Analysis of one-step and two-step real-time RT-PCR using SuperScript III.

Real-time reverse transcription polymerase chain reaction (RT-PCR) is a commonly used technique to analyze gene expression. There has been little research conducted to test if SuperScript III quantitative one-step (reverse transcription carried out in the same tube as PCR) and two-step (reverse transcription carried out in a separate reaction) RT-PCR systems provide similar real-time results. I...

One step forward

One-step and two-step bait fishing: Evaluation of results

The evaluation of the SILAC AP-MS experiments required a threshold for the SILAC ratio to define when an identified protein should be considered a contaminant or putative interaction partner. Other studies apply a statistical measure such as the Grubbs outlier test [1] or the z -test [2] to measure whether the deviation of a SILAC ratio is significant. These tests presuppose that the parameter ...