GeneReleaser

Details for Product No. ABIN3188319, Supplier: Log in to see
Application
DNA Extraction (DEx), Sample Preparation (Prep)
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Purpose GeneReleaser is a reagent, which releases DNA from whole blood, cell cultures, bacterial colonies etc.
Characteristics
  • DNA preparation before primer extension in 10 minutes
  • Facilitates the release of DNA from whole blood, tissues, bacteria, yeasts and plants
  • Sequesters cell lysis products which might inhibit thermostable polymerases or interfere with primer extension
  • Application Notes GeneReleaser® is a proprietary reagent which releases DNA from whole blood, cell cultures, bacterial colonies and the like. Lysis is accomplished directly in the amplification tube on a thermocycler. Typically, 1 μL of specimen is lysed in a total volume of 20 μL and amplification reagents are subsequently added to initiate amplification. GeneReleaser® sequesters cell lysis products which might inhibit polymerases and improves amplification yield and specificity. GeneReleaser® greatly simplifies the amplification of genomic DNA by avoiding the requirement to purify DNA. This avoids lengthy protocols and excessive sample manipulations which may introduce contamination.
    Protocol GeneReleaser is a reagent, which releases DNA from whole blood, cell cultures, bacterial colonies etc.
    Lysis is accomplished directly in the amplification tube on a thermocycler.
    Typically, 1 μL of sample is lysed in a total volume of 20 μL and amplification reagents are subsequently added to initiate amplification.
    GeneReleaser sequesters cell lysis products which might inhibit polymerases and improves amplification yield and specificity.
    GeneReleaser greatly simplifies the amplification of genomic DNA by avoiding the need to purify DNA.
    This avoids lengthy protocols and excessive sample manipulations, which may introduce contamination.
    GeneReleaser is provided as a ready-to-use stock sufficient for 100 or 400 assays.
    Assay Procedure

    General:
    It is assumed that the user is familiar with the programming and use of thermocyclers and the reactions which can be performed on them. No license to use PCR* is given or to be implied by this product or these instructions. A limited license for PCR can be obtained with the purchase of PCR reagents or thermocycling devices from Perkin Elmer.

    Whole Blood Procdure:

    1. Place 1 μL of whole blood into the bottom of each thermocycling tube for each specimen.
    2. Thoroughly resuspend the contents of the GeneReleaser® tube by inverting 10- 20X.
    3. Add 20 μL of the resuspended GeneReleaser® to each tube.
    4. . Perform the thermocycle program described below.
    5. Perform the amplification reaction according to your optimized protocol.
    Bacterial Colony Procedure:
    1. Place a single well isolated colony into the bottom of a thermocycle tube for each specimen.
    2. Perform steps 2-5 as per these steps in the Whole Blood Procedure above.
    Tissue Culture Procedure:
    1. Place 1 μL of cells at a density of 103-108 cells/mL into the bottom of a thermocycle tube for each specimen.
    2. Perform steps 2-5 as per these steps in the Whole Blood Procedure above.

    Thermocycle Program: NOTE: Reaction volumes and concentrations should be adjusted proportionally to allow for the 20 μL GeneReleaser® volume. In some instances a new magnesium titration may need to be performed to assure optimum amplification. We suggest the range of 1.5 mM - 4.0 mM Mg in 0.5 mM increments for this titration.

    Overlay the specimens with mineral oil.
    1. 65 °C hold 30 sec.
    2. 8 °C hold 30 sec
    3. 65 °C hold 90 sec.
    4. 97 °C hold 180 sec.
    5. 8 °C hold 60 sec.
    6. 65 °C hold 180 sec.
    7. 97 °C hold 60 sec.
    8. 65 °C hold 60 sec.
    9. 80 °C hold.
    10. Adjust temperature as necessary.
    11. Add amplification reagents without vortexing or otherwise mixing the amplification tube contents.
    12. Begin your optimized amplification. NOTE: It is very important that the very first denaturing step of the first cycle be at 94 °C for 2-5 minutes depending on brand of cycler, reaction volumes etc.

    Restrictions For Research Use only
    Storage 4 °C
    Storage Comment 4°C. Do not freeze.