T7 RNA Synthesis Kit

Introduction T7 RNA Synthesis Kit is designed for in vitro transcription of large amounts of RNA, provides with T7 RNA polymerase mix and four individually available nucleotides. The kit can also be used to synthesise capped RNA by adding cap analogues; or to synthesise modified RNA by replacing with modified UTP according to experimental requirements. The kit contains LiCl Solution for removing free nucleotides, enzymes and most of the template from the transcription product to obtain coarsely purified RNA.

Advantage

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 various RNAs, including but not limited to:

mRNA for vaccine or gene expression
precursors of circular RNA
labeled RNA probes
Modified RNA: prepared by admixing modified NTP (e.g., aminoallyl-, biotin-, fluorescein-, digoxin-NTP)

Performance

RNA yield

Fig. 1 Yield for different reaction durations

1 μg of 2600 bp DNA template was added to 20 μL of the reaction system and incubated at 37 °C for 30~120 min. The IVT products were purified with LiCl and analysed by NanoDrop™ to determine the amount of product for different reaction durations (Figure 1.A); and an aliquot of the product was electrophoresed on a 1 % agarose gel (Figure 1.B).

Competitor Comparison

Fig. 2 Comparison of in vitro transcribed RNA yields

Compare the yield of RNA for in vitro transcription with PC028+, PC035 kit, and a competitor IVT kit (NEB, #E2040). PC028+ represents the substitution of GeneCopoeia’s T7 RNA polymerase for the T7 RNA polymerase in the competitor’s IVT kit. The reaction system was configured using 1 μg of Control Template DNA from the PC035 kit as a template and incubated at 37 degrees for 2 hours. The final products were purified by LiCl and 1 μL of each was taken for electrophoretic analysis (Figure A). And Compared the yields calculated after measuring the concentration using Nanodrop (Figure B). The ssRNA ladder was loaded with 2 μL.

FAQs

1.Low transcript yield or failure

Use the Control DNA Template in the kit as a control template. Please contact us if control yields are low. If the experimental yield is normal in the control group but low in the experimental group, you can try:

Repurify the template to determine the purity of the template (260/230>2, preferably >2.2, and also 260/280>1.8), and electrophoresis of the product as a single band to confirm that the linearisation is complete.
Prolong the reaction time at 37℃, and increase the template input.
Configure more than one reaction system or expand the reaction system to improve the yield. The reaction time was extended at 37℃ and the amount of template input was increased.

2.Low yield of short transcripts

When the transcription product length is <300 nt, the reaction time can be extended to 4-16 h and the amount of DNA template can be increased to 2 μg in order to increase the yield of short transcripts.

3.Electrophoresis tailing of transcription products

The T7 RNA Polymerase Mix already contains RNase inhibitor, which can effectively inhibit product degradation. It is recommended to perform denaturing electrophoresis and use RNA loading buffer.

4. RNA transcription length is greater than expected

Products of higher than expected size in normal non-denaturing electrophoresis are generally RNA secondary structures, some of which cannot be removed even after simple heating. Please use denaturing electrophoresis and RNA loading buffer.

5.RNA transcription length is less than expected

Shorter-than-expected product lengths are generally a result of template problems, including transcription termination due to terminator-like secondary structures, etc., and require adjustment of the template sequence.

6.Transcription of Control DNA Template

Gluc with 5’UTR and Poly-A tail in the length of approximately 750 bp. The purified product can be used for cell transfection.

7.Why does the system yield less when scaled up?

It is recommended that the system be scaled up and distributed to small tubes for this purpose to avoid inhomogeneity of the reaction system during prolonged reactions, and also to re-mix the product every hour.

Related Products

Taq DNA Polymerase Kit (Cat. No. PC003)
Super Taq DNA Polymerase Kit (Cat. No. C0102A)
UltraHiPF® DNA Polymerase Kit
Clone Constrction for IVT (please inquire)
RNaseLock™ RNase Inhibitor (Cat. No. PC005)
DNase I
T7 RNA Co-transcription Synthesis Kit Coming Soon!
circRNA Synthesis Service (please inquire)

Price:	$260.00 $442.00
Catalog#:	PC035 PC036
XX:	50 Reactions 100 Reactions
Size:	50 Reactions 100 Reactions	T7 RNA Synthesis Kit – 20% Off* Ends April 30, 2025
Qty:
Manual:	Download Download

T7 RNA Synthesis Kit T7 RNA Synthesis Kit

*Promotions are valid in the US & Canada only. For international customers, please contact your local distributors. Promotion ends April 30, 2025. Discounts are not valid on previous purchases and cannot be combined with additional discounts.

Advantage

Application

RNA yield

Competitor Comparison