AmpliScribe™ SP6 High Yield Transcription Kit

Catalogue number

Pack size

AS3106

50 Reactions

AmpliScribe™ T3 High Yield Transcription Kit

Catalogue number

Pack size

AS3103

50 Reactions

AmpliScribe™ T7 High Yield Transcription Kit

Catalogue number

Pack size

AS2607

25 Reactions

AS3107

50 Reactions

AmpliScribe™ T7, T3 and SP6 High Yield Transcription Kits are specially formulated to utilise high concentrations of NTPs that are inhibitory to other in vitro transcription systems. AmpliScribe™ High Yield Transcription reactions can convert up to 90% of input NTPs into RNA, yielding >20-fold more full-length RNA transcript than conventional in vitro transcription reactions. AmpliScribe™ reactions consistently produce the highest yield of RNA from an in vitro transcription reaction. The RNA produced is full length, and the kit is specially formulated and quality tested to assure RNA integrity.(Figure 1)

Figure 1. The AmpliScribe™ T7 High Yield Transcription Kit is specially formulated to consistently produce >20-fold more RNA than a conventional T7 in vitro transcription reaction.

Notes

1. Use AmpliCap™ High Yield Message Maker Kits for efficient production of RNA with an m7G[5']ppp[5']G cap structure on the 5'-end.
2. Because of the very high concentration of NTPs used in AmpliCap™ reactions, these kits are not recommended for preparing high specific activity radiolabelled RNA probes. To generate high specific activity RNA probes, use the RiboScribe™ RNA Probe Synthesis Kits.
3. Use the DuraScribe™ T7 Transcription Kit to produce high yields of DuraScript™ RNA that is completely resistant to RNase A digestion.

Last modified: 2008-05-14 16:46:46

Applications

• Generation of RNA from DNA cloned downstream from a phage T7, T3 or SP6 RNA polymerase promoter
• Production of RNA for microinjection, RNAi (RNA Interference), anti-sense experiments and for RNA structural, splicing and processing studies
• Functional studies of heterogeneous nuclear RNAs, tRNAs, viral RNAs and ribozymes
• Produce fluorescent, biotinylated or digoxigenin -labelled RNA for use in microarray or in situ hybridisation
• Production of RNA for experiments using gene or expression arrays

Benefits

• High Yields - AmpliScribe™ reactions consistently incorporate up to 90% of the high level of NTPs in the reaction into RNA to produce the highest possible yields of full length RNA. A standard 20µl reaction with 1µg of DNA template yields up to 150µg of RNA in two hours using AmpliScribe™ T7 or AmpliScribe™ T3 RNA Polymerase and up to 100µg RNA using AmpliScribe™ SP6 RNA Polymerase
• High Yields for all template sises - synthesise both long (figure 2)and short (Figure 3)RNA using only one kit
• Scale up an AmpliScribe High Yield reaction to produce milligram amounts of RNA (figure 4)
• Produce microgram amounts of RNA from as little as 1ng of template (Figure 5)
• Produce >20 times more RNA than conventional or 'homemade' in vitro transcription reactions
• RNase inhibitor is included in the Enzyme Solution to ensure integrity of the RNA product
• AmpliScribe Kits are the best value for in vitro transcription

Figure 2. An AmpliScribe™ T7 High Yield Transcription reaction produces full-length, long RNA transcripts. Aliquots from standard 20-µl AmpliScribe T7 High Yield Transcription reactions were loaded onto a 1% agarose-formaldehyde gel and stained with ethidium bromide. The size of the RNA transcripts and RNA yields (in parentheses) were: Lane 1, 3.0 kb (185 µg); Lane 2, 4.0 kb (178 µg); Lane 3, 5.0 kb (176 µg); Lane 4, 6.0 kb (169 µg); M, RNA size marker

Figure 3. An AmpliScribe™ T7 High Yield Transcription reaction produces full-length, short RNA transcripts. Aliquots from standard 20-µl AmpliScribe T7 High Yield Transcription reactions were loaded onto a 12% polyacrylamide gel and stained with ethidium bromide. The size of the RNA transcripts and RNA yields (in parentheses) were: Lane 1, 46 bases (41 µg); Lane 2, 69 bases (55 µg); Lane 3, 88 bases (61 µg); Lane 4, 95 bases (65 µg); Lane 5, 242 bases (78 µg).

Figure 4. The standard 20-µl AmpliScribe™ T7 High Yield Transcription reaction can be scaled-up to produce milligram amounts of full-length RNA.

Figure 5. Limiting amounts of DNA template is efficiently transcribed in an AmpliScribe™ T7 reaction.

Last modified: 2008-05-14 16:46:46

Protocols for: AmpliScribe™ T7, T3 & SP6 Transcription Kits

AmpliScribe™ T7, T3 & SP6 Transcription Kits Protocol

(catalogue number AS2606 / AS3106 / AS2603 / AS3103 / AS2607 / AS3107)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Johnson, M.G. (1995) EPICENTRE Forum 2 (5), 4.
2. DeLong, E.F. et al. (1999) App. and Environ. Microbiol. 65, 5554.
3. Kaplan, E.D. et al. (1996) EPICENTRE Forum 3 (2), 1.
4. Hoffman, L.M. and Johnson, M.G. (1994) BioTechniques 17, 372.
5. Schanke, J.T. (2000) EPICENTRE Forum 7 (2), 6.

Last modified: 2008-05-14 16:46:46