RT-qPCR Extraction Control Orange

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Catalogue No: abx461077
Price: US$2,769.50
(Size: 500 rxns)

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RT-qPCR Extraction Control Orange

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Datasheet SDS
RT-qPCR Extraction Control Orange (emission wavelength 560 nm). This product consists of artificial cells containing an internal control RNA sequence with no known homology to any organism, and a specific primer/probe control mix for PCR detection. It can be used to monitor the RNA extraction process in RT-qPCR assays.

Target RT-qPCR Extraction Control Orange
Tested Applications PCR
Form Liquid
Storage Store at -80 °C. Avoid repeated freeze/thaw cycles.
Validity Up to 12 months.
Buffer The exact formulation is proprietary.
Kit Components
Component Volume
Internal Control RNA 5 × 200 µl
25X Control Mix 5 × 100 µl
50 mM MgCl2 1.2 ml
Availability Shipped within 10-20 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use All reactions should be carried out at room temperature unless stated otherwise.

Extraction Steps:
  1. Spin down all tubes before opening.
  2. Add 2 µl Internal Control RNA into each sample.
  3. Follow the manufacturer's protocol for total RNA extraction.
  4. Elute total RNA to a volume of 100 µl.
  5. Use 5 µl of the elution volume for a 20 µl reaction volume.

Recommended reagent volumes per 20 µl qPCR mix:

Component Volume
RT-qPCR Mix with 3 mM MgCl2 (not provided) 10 µl
Target Probe/Primer Mix (not provided) Variable
Extracted RNA Template (not provided) Variable
25X Control Mix
Vortex the Control Mix tube before use		 0.8 µl
50 mM MgCl2 1.2 µl
Reverse Transcriptase (not provided) 0.2 µl
RNase Inhibitor (not provided) 0.4 µl
Total Volume 20 µl

Assay Setup:

Step Number of Cycles Temperature Time per Cycle
Reverse Transcription 1 cycle 42 °C 10-20 min
Activation 1 cycle 95 °C 3 min
Denaturation30-40 cycles 95 °C 10 seconds
Annealing/Extension/Acquistion60 °C 30-45 seconds

Notes:
  • The fluorescence signal for the Internal Control RNA can be observed at 560 nm.
  • It is recommended that the end user carries out a validation step to ensure that the primers used do not cross-react with the Internal Control RNA.
  • Ct values of the Internal Control may vary due to elution volumes of nucleic acid and reagents used.
  • The optimal conditions will vary from reaction to reaction and are dependent on the system used.
  • Each parameter needs to be adjusted by the end user and some optimization may be required. For example, the standard annealing temperature is 60 °C, though this may need to be optimized by the user depending on the RT-qPCR mix used.
Research Articles on RT-qPCR Extraction Control Orange


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