ACCURIS inSTRUmEnTS qMAX Green qPCR Mix with Blue Tracking Dye Instructions
- June 12, 2024
- ACCURIS INSTRUMENTS
Table of Contents
ACCURIS inSTRUmEnTS qMAX Green qPCR Mix with Blue Tracking Dye Instructions
Description
The qMAX Green qPCR Mix, with blue tracking dyeis a ready-to-use, ultra-stable 2x master mix for use in real-time quantitative PCR assays with the option of post amplification melt profiles. The 2x master mix contains a non-inhibitory intercalating dye, HS Taq DNA Polymerase, and an optimized buffer with an inert blue tracking dye for easy visualization when pipetting. The mix provides robust realtime PCR with earlier cycle values (Ct) and broad range detection for increased sensitivity, speed, and room temperature stability. Ideal for genomic DNA, cDNA, & dilute templates.
- Ideal for fluorescent DNA/cDNA detection, gene expression analysis and sequence variant screening.
- Utilizes high quality, Accuris Hot Start Taq Polymerase to reduce nonspecific binding and provide easy reaction set up.
- A unique combination of salts, pH and PCR enhancers allow for detection of dilute targets and earlier Ct values.
- Contains an inert blue trackingdye to facilitate pipetting/visualization in s plates.
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. Accuris qMAX 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 9001:2015 standards.
Reaction setup
Briefly vortex the 2X mix before adding to the reaction
| Component | 20 µl reaction | Final concentration |
|---|---|---|
| Accuris qMAX 2X Green Master Mix | 10 µl | 1X |
| Forward Primer (10µM) | 0.8 µl | 400 nM |
| Reverse Primer (10µM) | 0.8 µl | 400 nM |
| Template DNA | 100 ng cDNA, 1 µg genomic | Variable |
| PCR-grade water | Up to 20µl 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 SYBR Green or FAM channel.
PCR Program
| Step | Temperature | Time |
|---|---|---|
| Initial denaturation | 95°C | 2-5min. (3+ min. for gemonic DNA) |
| 40 cycles* | 95°C | 5 seconds |
| 60° – 65°C | 10-40 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 application.
Package contents and reordering
Accuris qMAX Green qPCR Mix, with blue tracking dye is supplied in 100, 500
and 1000 reaction (20µl) packages.
Accuris qMAX Green qPCR Master Mix, with Blue tracking dye – Sample (20
rxns)
Catalog number: Low ROX – PR2000-L-S
Catalog number: Low ROX – PR2000-L-S
Includes 200µl of 2X Master Mix (20 rxns)
Accuris qMAX Green qPCR Master Mix, with Blue tracking dye, 100 rxns
Catalog number: Low ROX – PR2000-L-100
Includes 1.0ml of 2X Master Mix (100 rxns)
Accuris qMAX Green qPCR Master Mix, with Blue tracking dye, 500 rxns
Catalog number: Low ROX – PR2000-L-500
Includes 5×1.0ml of 2X Master Mix (500 rxns)
Accuris qMAX Green qPCR Master Mix, with Blue tracking dye, 1000 rxns
Catalog number: Low ROX – PR2000-L-1000
Includes 10×1.0ml of 2X Master Mix (1000 rxns)
- Use -N/L/H to indicate ROX requirements (ex: PR2000- L-100 = Low ROX mix*
General Guidelines
2X Taq Master Mix
The Master Mix contains HS Hot Start DNA polymerase, a proprietary,
fluorescent binding dye, dNTPs and an optimized buffer designed specifically
for maximum efficiency, sensitivity and successful quantitative PCR.
Amplicon
The optimal amplicon length should from 80 to 200 base pairs. Length should
not exceed 400 base pairs.
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/).
Reference Dyes (ROX™)
ROX passive reference dyes are required by some real-time PCR instruments. Not
all instruments 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.
Technical Support
For trouble-shooting and tech support, contact us by phone at 908 769-5555 or email [email protected]. When possible, please include instrument model, reaction conditions, PCR parameters, amplicon size and any traces and melting profiles.