ACCURIS PR2001 qMax Probe High ROX qPCR Mix Reactions Instructions
- June 13, 2024
- ACCURIS
Table of Contents
Accuris™ qMax™ Probe High ROX qPCR Mix
Description
Accuris qMax Probe qPCR Mix is a single tube formulation for sensitive and
efficient real-time, quantitative PCR assays utilizing probe detection
technologies, including
TaqMan®, Scorpions® and molecular beacon probes. The mix is optimized for
earlier threshold detection cycles (Ct ), fast cycling with exceptional,
reproducible results and low PCR inhibition.
Refer to www.accuris-usa.com/PCRReagents to determine the ROX level appropriate for your specific
cycler.
-Ideal for multiplex qPCR, two step RT-PCR, gene expression analysis, probe based detection of DNA/cDNA and screening of sequence variants.
-Utilizes high quality, Accuris Hot Start Taq Polymerase to reduce the formation of primer-dimers and provide easy reaction set up on the bench.
-Unique buffer formulation works for both single and multiplex qPCR.
-Compatible with both standard and fast cycling protocols.
Storage
Upon receipt, immediately store at -20°C.
Avoid excessive freeze/thaw cycles. When stored as directed, this product will
retain its activity for 12 months from date of receipt. The product may also
be stored at 4°C for up to one month.
Limitations of Use
For research purposes only. Not intended for therapeutic or diagnostic use.
Quality Control
Accuris qPCR mixes are tested for efficiency, activity, sensitivity,
processivity, heat activation, and absence of nuclease and nucleic acid
contamination. This product is manufactured under a comprehensive quality
management system, following ISO 9001:2008 standards.
General Guidelines
1. 2X Taq Master Mix
The Master Mix contains Accuris Taq Hot Start DNA polymerase, dNTPs and an
optimized buffer designed specifically for maximum efficiency, sensitivity and
successful quantitative PCR using TaqMan probes.
2. Amplicon
The optimal amplicon length should from 80 to 200 base pairs. Length should
not exceed 400 base pairs.
3. Primers
Primers should have a predicted melting temperature (Tm) of approximately
60°C, using primer design software such as Primer 3
(http://frodo.wi.mit.edu/primer3) or visual OMPTM
(http://dnasoftware.com/). Probe Tm should be 6° – 10°C higher than that of
the primers. For TaqMan® probes, avoid terminal guanosine residues by choosing
a probe close to the 5’ primer.
4. Reference Dyes (ROX™)
ROX passive reference dyes are required by some real-time PCR instruments.
Not allinstruments require the same level of ROX, and many of the newer
instruments do not require passive reference but include the option to use it
for normalization. To determine which kit matches the ROX level required by
your instrument, visit www.accuris-usa.com/PCR-Reagents.
Comparisons between suppliers should always been done in a 10-fold
amplification series. Low concentration loss of detection is the only direct
measurement of sensitivity.
Technical Support
For trouble-shooting and tech support, contact us by phone at 908 769-5555 or
email info@accuris-usa.com. When possible,
please include instrument model, reaction conditions, PCR parameters, amplicon
size and any traces and melting profiles.
Accuris is not responsible for consequential or incidental damages, whether
direct or indirect, resulting from use of this product.
Reaction setup
Briefly vortex the 2X mix before adding to the reaction
| Component | 20 µl reaction | Final concentration |
|---|---|---|
| Accuris qMax Probe Master Mix | 10 µl | 1X |
| Forward Primer (10µM) | 0.8 µl | 400 nM |
| Reverse Primer (10µM) | 0.8 µl | 400 nM |
| Probe (10µm) | 0.4µl | 200 nM |
| Template DNA | <100 ng cDNA, <1 µg genomic | variable |
| PCR-grade water | to final reaction volume |
For other volumes, adjust the amount of each component accordingly.
Gently mix the solution. If needed, spin briefly in a microcentrifuge to bring
reaction mixture to the bottom of the tube. Transfer samples to a real time
thermal cycler, acquiring data on the appropriate channel.
PCR Program
| Step | Temperature | Time |
|---|---|---|
| Initial denaturation | 95°C | 2 minutes (3 minutes for genomic DNA) |
| 40 cycles* | 95°C | |
| 60° – 65°C | 5 seconds |
20-30 seconds
Melt Anaylsis (optional)
*Do not use temperatures below 60º or exceed 30 seconds.
Accuris guarantees the performance of this product as described when used in
accordance with these instructions. It is the responsibility of the purchaser
to determine the suitability of this product for their particular applicaiton.
Package contents and reordering
Accuris qMax Probe qPCR Master Mix, supplied in 100, 500 and 1000 reaction
(20µl) packages.
Accuris qMax Probe qPCR Master Mix, Sample
Catalog number: Low ROX – PR2001-L-S
High ROX – PR2001-H-S
No ROX – PR2001-N-S
Includes 200µl of 2X Master Mix (20 rxns)
Accuris qMax Probe qPCR Master Mix, 100 rxns
Catalog number: Lo ROX – PR2001-L-100
High ROX – PR2001-H-100
No ROX – PR2001-L-N-100
Includes 1.0ml of 2X Master Mix (100 rxns)
Accuris qMax Probe qPCR Master Mix, 500 rxns
Catalog number: Lo ROX – PR2001-L-500
High ROX – PR2001-H-500
No ROX – PR2001-N-500
Includes 5×1.0ml of 2X Master Mix (500 rxns)
Accuris qMax Probe qPCR Master Mix, 1000rxns
Catalog number: Lo ROX – PR2001-L-1000
High ROX PR2001-H-1000
No Rox – PR2001-N-1000
Includes 10×1.0ml of 2X Master Mix (1000 rxns)
One Tube Formulation, 2X Concentration
Package contains :
1.0m1 of 2X qMax Probe qPCR Mix 100 reactions, Based on 200 total reaction
volume
5.0m1 of 2X qMax Probe qPCR Mix 500 reactions, Based on 200 total reaction
volume
Store at -20°C upon receipt
PH: 908.769.5555
EM: Info@accuris-usa.com