T7 RNA Polymerase

Product Information

T7 RNA Polymerase is a DNA-dependent RNA polymerase with strict specificity for T7 promoter. Use double-stranded DNA (linearised plasmid or PCR product) as a template to synthesise RNA that is complementary to the reverse single-stranded DNA downstream of the promoter. T7 RNA Polymerase accepts modified nucleotides (e.g., biotin-, digoxigenin-, fluorescein-labeled nucleotides) as substrates for RNA synthesis.

Catalog#	Product	Size
PC028	T7 RNA Polymerase (50 U/μL)	100 μL
PC028	10× Transcription Buffer	250 μL
PC029	T7 RNA Polymerase (50 U/μL)	5× 100 μL
PC029	10× Transcription Buffer	5× 250 μL
Inquire	T7 RNA Polymerase ( >200 U/μL)

*For T7 RNA polymerase (high concentration, >200 U/μL) products, please contact us at 1-301-762-0888 or inquiry@genecopoeia.com.

Storage buffer: 50 mM Tris-HCl，100 mM NaCl，20 mM β-ME，1 mM EDTA，50% Glycerol，0.1% (w/v) Triton® X-100 (pH 7.9 @ 25°C)

Definition of activity unit: One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 μl in 1 hour at 37°C.

Advantages

Yields 150~200 μg of RNA from 1 μg of DNA template.
Optimized transcription buffer: Requires less T7 RNA polymerase for maximum RNA yield compared to other competitors.

Application

Synthesis of large amounts of RNA, including:

mRNA for vaccines or gene expression
circRNA precursors
Modified RNA: prepared by admixing modified NTP (e.g., aminoallyl-, biotin-, fluorescein-, digoxin-NTP)
labelled RNA probes, etc.

Performance Data

T7 RNA Polymerase reaction time course

Time course of RNA synthesis from a 1 μg 800 bp template.

Figure 1. Time course of standard RNA synthesis from 1 µg of a 2600 bp DNA template. Reactions were incubated at 37°C for different time in a PCR machine. The IVT products were purified by LiCl and (A) quantified on NanoDrop™ Spectrophotometer, and (B) equal volumes were loaded to the 1 % agarose gel for electrophoresis.

Transcription products in different length

Agarose gel analysis of different sized RNA generated using T7 RNA Polymerase

Figure 2. The in vitro transcription reactions were carried out by adding 250 ng of different sized template to a 20 µL reaction. As shown, the T7 RNA Polymerase is suitable for in vitro transcription reactions with different product lengths.

FAQs

T7 promoter sequence

5′-TAATACGACTCACTATAG-3′

T7 RNA polymerase starts transcription from the underlined G in the promoter sequence. Using the reverse strand as a template, transcription begins at 5′->3′. The first base in the transcript will be G.

Template for in vitro transcription with available T7 RNA polymerase?

Double-stranded DNA, such as linear plasmids or PCR products.The DNA template needs to be linearized with an enzyme that produces 5′ protruding sticky ends or blunt ends, or it can be linearized by PCR, with the amount of template required for the in vitro transcription reaction being reduced when linearised by PCR.

Reasons for heterozygous bands in transcription products?

First, make sure that the DNA is fully linearized as required and that the system is free from RNase contamination. If the IVT product is taken and directly subjected to normal electrophoresis, most of the spurious bands are formed due to the secondary structure of the RNA, which can be eliminated by denaturing at 70℃ for 1 min after the addition of RNA loading buffer. Also, there are a small number of unavoidable dsRNA by-products that can be inhibited by adding 1M urea to the reaction, or removed by subsequent purification.

Price:	$70.00 $280.00
Catalog#:	PC028 PC029
XX:	5,000 U 25,000 U
Size:	5,000 U 25,000 U
Qty:
Manual:	Download Download

T7 RNA Polymerase T7 RNA Polymerase

Application

T7 RNA Polymerase reaction time course

Transcription products in different length

Figure 2. The in vitro transcription reactions were carried out by adding 250 ng of different sized template to a 20 µL reaction. As shown, the T7 RNA Polymerase is suitable for in vitro transcription reactions with different product lengths.

5′-TAATACGACTCACTATAG-3′

T7 RNA polymerase starts transcription from the underlined G in the promoter sequence. Using the reverse strand as a template, transcription begins at 5′->3′. The first base in the transcript will be G.

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