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Random priming is widely used in 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:
RNA without poly(A) tail such as ribosomal RNAs.
RNA with strong secondary structures.
Partially degraded RNA samples.
Target regions at 5´ end of a long messenger RNA transcript.
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.
Reagents Supplied
The following reagents are supplied with this product:
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.
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.)
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
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]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
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.)
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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