The construction was done by polymerase chain reaction (PCR -based mutagenesis involving two sequential stePCR -basedt al. 2004).
The two homologous arms for SirT1 and HDAC2 KO vector construction were obtained by polymerase chain reaction (PCR) and purified using gel extraction kits.
Tissue samples obtained before heat-shock (BHS), at the cessation of heat-shock (CS), and 3, 12, and 24 hours after the cessation of heat-shock (ACS), were used for reciprocal SSH library construction and quantitative reverse transcription - polymerase chain reaction (QPCR) analysis of gene expression using samples from a group that was transferred but not heat-shocked (CT) as controls.
We describe methodologies for the rapid, efficient, and economical construction of such libraries using inverse polymerase chain reaction (PCR).
All mutagenic polymerase chain reaction (PCR) primers utilized in the construction of PfTR mutants were purchased from Integrated DNA Technologies Coralville, IAA).
A novel tri-primer polymerase chain reaction method (TP-PCR) was developed for the construction of a fused fpg gene, in which no endonuclease and ligase were used.
Additional details regarding cell culture, RNA isolation, small RNA cDNA library construction, massively parallel sequencing, computational pipeline description and miRNA TaqMan quantitative reverse-transcription polymerase chain reaction (qRT-PCR) are provided in detail in Supplementary Methods S1 and in [23].
Polymerase chain reaction-based markers have been widely used in the construction of genetic linkage maps for B. oleracea [ 13, 14], B. nigra [ 15], B. juncea [ 16], and B. napus [ 17, 18].
The plastid DNA is sometimes enriched by rolling circle amplification, long-range polymerase chain reaction (PCR), and bacterial artificial chromosome or fosmid library constructions.
Polymerase chain reaction (PCR) was performed with Phusion DNA polymerase (New England Biolabs) for construction of the vectors and sequencing purposes and ExTaq (Takara) or Taq (NEB) for diagnostic purposes.
