genesig Real-time PCR Internal Controls: RNA Internal Extraction Control (Cy5 Probe)

Kits and Assays / Kits-PCR, genesig™

-20°C

Real-time PCR Internal Controls:
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

About This Product:
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.

RNA Internal Extraction Control Kits
When performing RNA extraction, it is often advantageous to have an exogenous source of RNA template that is spiked into the lysis buffer. This control RNA is then co-purified with the sample RNA 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 RNA also indicates that PCR inhibitors are not present at a high concentration.

A separate primer and probe mix are supplied with this kit to detect the exogenous RNA following cDNA synthesis using real-time PCR. The primers are present at PCR limiting concentrations which allows detection of the control cDNA and target cDNA in a multiplexing reaction. Amplification of the control cDNA 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 cDNA. A dye should be selected that reads through a separate channel to the target gene.

ChannelExc WavelengthEm Wavelength
FAM495520
Cy5649670
TAMRA555576
VIC530549

The exogenous RNA 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.

Kit Contents:
• Lyophilised Internal control template RNA (150 reactions BLUE)
• Lyophilised Internal control primer/probe mix (150 reactions BROWN)
• Internal Control RNA RT primer mix (150 reaction GREEN)
• RNAse/DNAse free water (WHITE)

User-Supplied Reagents and Equipment:
• 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