• My NEB
  • Print
  • PDF
    •

    Random Primer Mix

    Description

    Random priming is widely used in the first strand cDNA synthesis. Random Primer Mix is a ready-to-use optimized mixture of hexamers and anchored-dT primer (dT23VN). A mixture of hexamers and anchored-dT primer provides even and consistent coverage of the RNA template population across a wide range of RNA template concentration. In contrast, traditional random priming by hexamer has poor coverage of 3´ end of RNA templates (1,2,3).

    We recommend using Random Primer Mix for reverse transcription of the following RNA templates:

    1. RNA without poly(A) tail such as ribosomal RNAs.
    2. RNA with strong secondary structures.
    3. Partially degraded RNA samples.
    4. Target regions at 5´ end of a long messenger RNA transcript.
    Figure 1: 
    Figure 1
    Various amounts of human spleen total RNA (2 μg to 20 pg) were reverse transcribed using ProtoScript First Strand cDNA Synthesis Master Mix Kit (NEB #E6300 ) in the presence of 6 μM Random Primer Mix (panel A) or 3.5 μM random hexamers (panel B). About 1/10th of the cDNA products were then analyzed in duplicate qPCR reactions using a pair of primers specific for the beta actin gene with DyNAmo™ FLASH SYBR® Green qPCR Master Mix on a Bio-Rad IQ5® system.

    Properties and Usage

    Storage Temperature

    -20°C

    Notes

    1. Recommended Usage: Random Primer Mix (60 µM) contains 35 µM hexamers and 25 µM dT23VN  and 1 mM dNTPs. We recommend using Random Primer Mix at a final concentration of 6 µM.

    References

    1. Goff, L.A. et al. (2004). BMC Genomics. 5, 76.
    2. Djikeng, A. et al. (2008). BMC Genomics. 9, 5.
    3. Rashtchian, A. (1994). Genome Research. 4, 83.

    FAQs

    1. What products does NEB offer for functional genomics and reverse genetics?

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

    Interactive Tools

    Certificate of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.