THE PROBLEM:
DNA-PCR and RNA-RT-PCR assays need to control for the following: Extraction of DNA or RNA.
Amplification.
DNA-PCR and RNA RT inhibition.
Reagent quality including dNTPs, Buffers, Taq, and reverse transcriptase.
Instrument function, including thermal cycling and fluorescent detection.
THE ANSWER:
The Attostar answer to these control questions is to use bacteriophage–T4 for DNA and Qß for RNA.
The Attostar regents include: T4 and Qß bacteriophage,
PCR master mixes,
Primers and probe reagents,
Plasmid DNA.
Detection of the bacteriophage internal control DNA/RNA signifies a successful extraction and amplification.
Failure to detect the bacteriophage internal control DNA/RNA means the extraction or amplification procedure failed.
To use these reagents, T4 or Qß is added directly to the sample. The sample is then extracted for DNA or RNA. The extracted nucleic acid, now containing T4 or Qß and the sample nucleic acids are used for PCR or RT-PCR.
The probe for real time detection of this T4 DNA or Qß RNA target is a molecular beacon1 containing a fluorophore such as FAM or Quasar 670 and a quencher such as BHQ1 or BHQ22.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
1: This product is sold under license from the Public Health Research Institute. It may be used under PHRI Patent Rights only for the purchaser’s research and development activities.
2: ‘Black Hole Quencher,’, ‘CAL Fluor,’ ‘Pulsar’ and ‘Quasar ’ are trademarks of and licensed by Biosearch Technologies, Inc., Novato, CA. The BHQ, CAL Fluor, Pulsar and Quasar dye technology is the subject of existing or pending patents including US Patent No. 7,019,129 and is licensed and sold under agreement with Biosearch Technologies, Inc.