ProtoScript® II First Strand cDNA Synthesis Kit

Catalog #	Concentration	Size	List Price	Quantity	Your Price
E6560L		150 reactions	$1,103.00		$992.70
E6560S		30 reactions	$275.00		$247.50

Catalog #	Size	List Price	Your Price
E6560L	150 reactions	$1,103.00	$992.70
E6560S	30 reactions	$275.00	$247.50

ProtoScript II First Strand cDNA Synthesis Kit features two optimized mixes, ProtoScript II Enzyme Mix and ProtoScript II Reaction Mix. The enzyme mix combines ProtoScript II Reverse Transcriptase and Murine RNase Inhibitor, while the reaction mix contains dNTPs and an optimized buffer. ProtoScript II Reverse Transcriptase is a recombinant M-MuLV reverse transcriptase with reduced RNase H activity and increased thermostability. It can be used to synthesize first strand cDNA at higher temperatures than the wild type M-MuLV. The enzyme is active up to 48°C, providing higher specificity and higher yield of cDNA.

The kit also provides two optimized primers for reverse transcription and nuclease-free water. An anchored Oligo-dT primer [d(T)₂₃VN] forces the primer to anneal to the beginning of the polyA tail. The optimized Random Primer Mix provides random and consistent priming sites covering the entire RNA templates including both mRNAs and non-polyadenylated RNAs. The first strand cDNA product generated is up to 10 kb.

Quantitative detection of reverse transcription products using the ProtoScript II First Strand cDNA — Ten-fold serial dilutions of in vitro transcribed luciferase RNA (1x10² to 1x10⁹ copies) were mixed with 1 ng Jurkat total RNA. The RNA was reverse transcribed using first strand cDNA synthesis kits, as indicated, and 1/10th of the cDNA product was used for qPCR using luciferase-specific primers and iTaqTM Universal SYBR Green Supermix (BioRad). Ct values were plotted as a function of input luciferase RNA equivalent copies. Each data point represents the mean +/- 95% confidence interval of a minimum of 5 amplification reactions. ProtoScript II – NEB ProtoScript II First Strand cDNA Synthesis Kit and random primer mix; SuperScript II – Invitrogen SuperScript First Strand Synthesis System for RT-PCR and random hexamers; SuperScript® III – Invitrogen SuperScript III First Strand Synthesis SuperMix and random hexamers.

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

E6560S -20

ProtoScript^® II Reaction Mix	M0555AVIAL	-20	1 x 0.4 ml	2 X
ProtoScript^® II Enzyme Mix	M0556AVIAL	-20	1 x 0.06 ml	10 X
Random Primer Mix	S1330AVIAL	-20	1 x 0.07 ml	60 µM
Nuclease-free Water	B1502AVIAL	-20	1 x 1.5 ml
Oligo d(T)₂₃ VN	S1332AVIAL	-20	1 x 0.07 ml	50 µM

E6560L -20

ProtoScript^® II Reaction Mix	M0555AVIAL	-20	5 x 0.4 ml	2 X
ProtoScript^® II Enzyme Mix	M0556AVIAL	-20	5 x 0.06 ml	10 X
Random Primer Mix	S1330AVIAL	-20	5 x 0.07 ml	60 µM
Nuclease-free Water	B1502AVIAL	-20	5 x 1.5 ml
Oligo d(T)₂₃ VN	S1332AVIAL	-20	5 x 0.07 ml	50 µM

Properties & Usage

Companion Products

References

Liao, J. and Gong, Z. (1997). Biotechniques. 23, 368-370.
Sambrook, J. and Russel, D.W. (2001). Molecular Cloning: A Laboratory Manual, (3rd ed.),. (pp. 8.46–8.53 avnd 11.37–11.42). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

FAQs

Citations

See more details on Bioz

Quality Control Assay

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.

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]

E6560S_L_v1

Certificate of Analysis

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]

Safety Data Sheets

Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@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.