Optimal positive control for DNA/RNA extraction and PCR conditions Unique product features • Perfect Internal control • Taqman style probe • Available to read through CY5, FAM, TAMRA, VIC or ROX channel. • Primer limited to prevent interference with target gene primers

Negative test results are only valid when the suitability of the reagents, template and protocols have also been demonstrated. The Primerdesign real-time PCR internal control kit contains either control RNA or DNA. This can be spiked in to your sample prior to nucleic acid extraction or diluted and spiked directly in to your real-time PCR reaction. A positive signal of around Cq 25 indicates that your experimental procedure is working well.

The internal control kit can be multiplexed, or used in parallel with your test samples. The assay is primer limited and based on an obscure nucleic acid sequence so will not therefore interfere with the detection of your target sequence.

DNA Internal Extraction Control Kits When performing DNA extraction, it is often advantageous to have an exogenous source of DNA template that is spiked into the lysis buffer. This control DNA is then co-purified with the sample DNA and can be detected as a positive control for the extraction process (internal extraction control). Successful co-purification and real-time PCR for the control DNA also indicates that PCR inhibitors are not present at a high concentration.

Alternatively, the DNA in this kit can be spiked directly into a PCR reaction to confirm directly that the test sample is competent to support real-time PCR amplification (internal PCR control).

A separate primer and probe mix are supplied with this kit to detect the exogenous DNA using real-time PCR. The primers are present at PCR limiting concentrations which allows multiplexing with the target sequence primers. Amplification of the control DNA does not interfere with detection of the target gene even when the target gene is present at low copy number. A number of different dyes are available so that a range of channels can be used to detect the control DNA. A dye should be selected that reads through a separate channel to the target gene.

Channel Exc Wavelength Em Wavelength FAM 495 520 Cy5 649 670 TAMRA 555 576 VIC 530 549

The exogenous DNA sequence supplied with this kit can be used with all human and animal tissues except rat and when used according to the protocols, gives a CT value of 26+/-3.

• Lyophilised Internal control template DNA (150 reactions BLUE) • Lyophilised Internal control primer/probe mix (150 reactions BROWN) • RNAse/DNAse free water (WHITE)

• Real-Time PCR instrument • Mastermix or Mastermix components This kit is designed to work well with all commercially available Mastermixes. However, we recommend the use of Primerdesign 2x PrecisionPLUSTM Mastermix. • Pipettors and Tips • Vortex and centrifuge • DNA template