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RT² qPCR Primer Assay for Mouse Mlh1: PPM04973C

Catalog Number Description
PPM04973C MutL homolog 1 (E. coli)  
Gene Symbol: UniGene #: Refseq Accession #:
Mlh1 Mm.486383 NM_026810
Please click here to view the Band Size and Reference Positions information.
The RefSeq Accession number refers to the representative sequence used to design the enclosed primers. These primers can also generate amplicons from the following splice variants:   XM_006511946 ;XM_006511947 ;XM_006511948 ;XM_006511949 ;XM_006511950 ;XM_006511951 ;XR_379848 ;XR_379849 

Description   Find Other Genes
The RT² qPCR Primer Assay is the most reliable SYBR® Green-based quantitative real-time PCR assay for gene expression analysis. Our experimentally verified design algorithm yields gene-specific qPCR assays characterized by uniform and high PCR efficiencies and standardized amplification conditions. Every RT² Primer Assay is subjected to rigorous experimental verification. Single product amplification of the correct size and high PCR efficiency are guaranteed when using the appropriate RT² qPCR Master Mixes. The uniform PCR amplification efficiencies and PCR conditions of the RT² qPCR Primer Assays provide an accurate and scalable solution for multiple gene expression analyses. Browse Primer Assays By Gene
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Useful Links
Pricing and Ordering shRNA Plasmid Brochure
User Manual White Paper Instrument Setup Guide
Technical Resources Slide Presentation Register for Web Seminar
Protocol: For Experienced Users

NOTE: Do not use DEPC treated H2O. Use high-quality, nuclease-free H2O. If you are not sure whether your RNase, DNase-free water has been DEPC treated, please check with the supplier.

  1. Briefly (10-15 seconds) spin down all reagents.
  2. For each 25-µl PCR, mix the following components in a PCR tube:
    12.5 µl RT² SYBR Green qPCR Master Mix
    10.5 µl H2O
    1.0  µl  RT² First Strand cDNA (template)
    1.0 µl gene-specific RT² qPCR Primers

    25.0 µl final volume
     
    • Note: If your cDNA template is not synthesized using an SABiosciences RT² First Strand Kit, 1 to 10 µl of cDNA (generated from up to 250 ng total RNA) may be used. Adjust the H2O amount for a final reaction volume of 25.0 µl.
    • Note: For improved ease and consistency, RT² qPCR Primer Assay-specific pre-mixes may be prepared. Scale up the following formula for the number of reactions to be performed within a single experiment (plus 0.1X excess volume to allow for multiple pipetting).
    Assay-Specific Pre-Mix
    12.5 µl RT² SYBR Green qPCR Master Mix
    10.5 µl H2O
    1.0 µl gene-specific RT² qPCR Primers

    24.0 µl final volume

    For each 25-µl PCR reaction,
    mix the following in a PCR tube

    24.0 µl Assay-Specific Pre-Mix
    1.0  µl  RT² First Strand cDNA (template)

    25.0 µl final volume
  3. Quickly centrifuge and place your tubes in your real-time thermal cycler.
  4. Enter and run the appropriate program for your real-time instrument.
  5. Use a two-step cycling program, for the following instrumentation:

    ABI: 5700, 7000, 7300, 7500, 7700, 7900HT, StepOnePlus

    BioRad: iCycler®, iQ5, MyIQ

    Stratagene: Mx3000p, Mx3005p, Mx4000p

    Eppendorf: Mastercycler® ep realplex

    Cycles Duration Temperature
    1 10 minutes1
    95ºC
    40 15 seconds
    95ºC
    1 minute2 60ºC

    Roche*: LightCycler 480® Cycles Duration Temperature
    1 10 minutes1
    95ºC
    45 15 seconds
    95ºC
    1 minute2 60ºC

    *Attention Roche LightCycler 480 Users: Adjust the ramp rate to 1ºC/sec. Please refer to the Instrument Setup Guide at http://sabiosciences.com/pcrarrayprotocolfiles.php for more information on other REQUIRED changes to settings for Melt Curve Acquisition.

    Use a three-step cycling program, for the following instrumentation:

    BioRad: CFX96, CFX384, Opticon, Opticon 2, Chromo 4 (MJ Research)

    Takara: TP-800

    All other instruments

    Cycles Duration Temperature
    1 10 minutes1
    95ºC
    45 15 seconds 95ºC
    30 to 40 seconds2,3 55ºC
    30 seconds 72ºC

    The 10-minute step at 95ºC is required to activate the HotStart DNA polymerase.
    Detect and record SYBR Green fluorescence from every well during the annealing step of each cycle.
    Different instruments need different lengths of time to detect the fluorescent signal. Choose the appropriate time for the annealing step (55ºC) for your instrument.

 

Materials Included / Packing List

Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt.

One gene-specific PCR primer assay (10 µM each primer, 2 primers for 200 PCR reactions)
One Certificate of Performance

Storage Conditions
RT² qPCR Primer Assay are shipped at ambient temperature. For long-term storage, store RT² qPCR Primer Assays at -20 ºC.

NOTICE TO PURCHASERS

  • This product is intended for research purposes only and is not intended for drug or diagnostic purposes or for human use. Use of kit components for reproduction of any primer pair mix, to modify kit components for resale or to use RT² qPCR Primer Assays to manufacture commercial products without written approval of SABiosciences Corporation is expressly prohibited.

LIMITED PRODUCT WARRANTY

  • This warranty limits our liability to the replacement of this product in the event the product fails to perform due to any manufacturing defect. SABiosciences Corporation makes no other warranties of any kind, expressed or implied, including without limitation, warranties of merchantability or fitness for a particular purpose. SABiosciences Corporation shall not be liable for any direct, indirect, consequential or incidental damages arising out of the use, the results of use or the inability to use this product.

TRADEMARKS

  • RT² qPCR, SABiosciences, and the SA logo are trademarks of SABiosciences Corporation. SYBR is a registered trademark of Molecular Probes.