| Description |
| The RT² Real-Time™ ROX PCR master mix contains all of the reagents and
buffers required for standard TaqMan® Gene Expression Assays in ABI
instrumentation: real-time PCR buffer, a high-performance HotStart DNA Taq
polymerase, nucleotides, and the ROX reference dye needed to normalize the
instruments' optics. Simply add the master mix to PCR tubes containing
template and a TaqMan® Primer & Probe Set. The chemically-modified
and tightly controlled HotStart enzyme uniquely provides more accurate PCR
results by preventing the amplification of primer dimers and other
non-specific products. This RT² Real-Time™ PCR master mix is best
suited for real-time PCR applications using TaqMan® Gene Expression
Assays in ABI instrumentation. |
| Materials Included /
Packing List |
|
Please check the kit components
immediately after you receive this package. SABiosciences is not responsible
for any missing items not reported within two (2) business days upon
receipt.
Contents:
PA-114: Twenty (20) tubes each containing 1.25 ml of the 2X solution
and enough for 100 standard 25-µl reactions
PA-014: Two (2) tubes each containing 1.25 ml of the 2X solution and
enough for 100 standard 25-µl reactions
Storage Conditions: The kit is shipped on dry ice or cold packs.
For long-term storage, keep at -20 ºC. If entire volume is not be used
all at once, divide into aliquots and store at -20 ºC. Avoid repeated
freezing and thawing.
Shelf Life: All reagents are stable for 6 months after receipt if
stored at the recommended temperature. |
| Brief Protocol |
- Ensure that you do not contaminate the 2X real-time PCR cocktail by
dividing into aliquots containing the amount of cocktail necessary for
the number of reactions you are preparing each day. The rest of the
cocktail should be kept in storage away from any sources of template
DNA.
- For each 25-µl PCR, mix the following components in a PCR tube:
- 12.5 µl RT² Real-Time™ ROX PCR Master Mix
- 10.25 µl ddH2O
- 1.0 µl of either undiluted or dilute template cDNA
- 1.25 µl 20X TaqMan® Primer & Probe Set
25.0 µl final volume
- Place tubes in real-time thermal cycler. Enter and run the following
program.
95 ºC, 10 min; 40 cycles of
(95 ºC, 15 sec; 60 ºC, 60 sec).
NOTE: The 10 min step at 95 ºC is required to activate
the HotStart Taq DNA polymerase.
- Otherwise, program the real-time thermal cycler to detect and record
the probe signal as described by the instrument and expression assay
manufacturer.
ABI® and TaqMan® are registered trademarks of Applied Biosystems.
Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser’s own internal research. No other patent rights are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
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