Bio-Rad has added the mouse genome to its PrimePCR quantitative real-time PCR assays.
Bio-Rad collaborated with Biogazelle to design, optimize, and experimentally validate the SYBR Green-based gene expression assays. PCR primer pairs for each mouse transcript were designed to follow where possible strict guidelines on maximum transcript coverage, minimal overlap with known SNPs, and spanning large introns, the company said.
In accordance with the minimum information for publication of quantitative real-time PCR experiments, or MIQE, guidelines, every assay was wet-lab validated following strict assay performance standards for amplification efficiency, specificity, sensitivity, and linear dynamic range. Assay specificity was confirmed using next-generation sequencing. This approach yielded validated assays for more than 90 percent of the mRNA transcriptome, Bio-Rad said.
Bio-Rad in August launched its portfolio of MIQE-compliant qPCR assays and panels, developed under a partnership with Biogazelle (PCR Insider, 8/30/2012).
Integrated DNA Technologies this week launched its PrimeTime qPCR assay plates for high-throughput qPCR analysis.
Primers and probes for 5'-nuclease and intercalating dye assays (e.g. SYBR) can now be directly ordered online in a 96-well plate format, eliminating the time-consuming transfer of primers and probes from reagent stocks, IDT said. The new assay plates are ideal for many applications that generate large quantities of data, such as validation of next-generation sequencing and microarray data, and high-throughput gene expression screening.
A range of primer and/or probe concentrations are available lyophilized within a 96-deep-well plate. Different dye-quencher combinations can also be ordered on the same plate, extending the range of assays that can be performed during a single run, IDT said.
New England Biolabs has launched the Q5 SDM reagent kit for site-directed mutagenesis.
Q5 SDM is capable of introducing long insertions and deletions, allowing a broader range of applications for the SDM technique. The kit can make standard single-base modifications, as well as novel manipulations, such as deletions or additions of any length, including the addition of nuclear localization signals to cloned proteins, the company said.
NEB adopted a non-overlapping primer design to allow for longer modifications. The most popular SDM methods use overlapping primers that limit the length of insertions or deletions, the company said. Meantime, NEB's approach enables exponential amplification and more efficient plasmid transformation, which facilitates the successful modification of highly repetitive or GC-rich sequences.
The company also released an SDM primer design tool called NEBaseChanger to support the Q5 SDM kit.