DuraScribe® T7 Transcription Kit
The DuraScribe® T7 Transcription Kit* is an in vitro transcription kit that produces RNA – called DuraScript® RNA – that is completely resistant to RNase A digestion.
The DuraScribe T7 Transcription Kit includes DuraScribe T7 RNA Polymerase, an enhanced formulation of the Y639F mutant1 of T7 RNA Polymerase that efficiently incorporates 2´-Fluorine-CTP (2´-F-dCTP), 2´-Fluorine-UTP (2´-F-dUTP), ATP and GTP into RNase-resistant RNA transcripts called DuraScript RNA (Figure 1). The DuraScribe T7 RNA Polymerase uses the same T7 promoters as the wild-type T7 RNA Polymerase.
Applications
DuraScript RNA has demonstrated advantages for RNA aptamer synthesis, and ribozyme, RNA interference (RNAi) and antisense RNA studies.
Benefits
- DuraScript RNA is resistant to RNase A and DNase
- High Yields of DuraScript RNA
- T7 in vitro transcription reations. DuraScribe T7 RNA Polymerase uses the same T7 promoters as the wild type T7 RNA Polymerase.
- Ideal for RNA aptamer studies. Numerous citations using the DuraScribe Kit with RNA aptamers
- SELEX-compatible. DuraScript RNA can be used in SELEX selection processes.
- Make long or short DuraScript RNA. Double-strand oligos, linearized plasmids, and PCR products with a T7 polymerase promoter can be transcribed.
References
- Sousa, R. and Padilla, R. (1995) EMBOJ. 14:18 4609-4621.
Table 1. Yield of DuraScript® RNA from a DuraScribe® Kit reaction. One microgram of a 3-Kb DNA template was linearized at different sites and then transcribed in a DuraScribe T7 Transcription Kit reaction for 4 hours. The yield of DuraScript RNA produced from each template is shown in micrograms (µg) and in picomoles (pmol).
Size of DuraScript RNA produced | DuraScript RNA Yield (µg) | DuraScript RNA Yield (pmol) |
2600 nts | 100 µg | 116 pmol |
1400 nts | 58 µg | 124 pmol |
330 nts | 18 µg | 164 pmol |
88 nts | 9 µg | 307 pmol |
Figure 1 (click to enlarge). The DuraScribe® T7 RNA Polymerase efficiently incorporates 2´-F-dCTP and 2´-F-dUTP into full-length DuraScript® RNA. The presence of the fluorine at the 2´-position of the 2´-F-dC and 2´-F-dU nucleotides prevents digestion by RNase A.
Figure 2. Yield of RNA from a DuraScribe® T7 Transcription reaction. A standard reaction (4-6 hours) produced 40-60 µg of a 1.4-kb DuraScript® RNA.
Figure 3. DuraScript® RNA is resistant to RNase A digestion. A 1.4-kb standard RNA transcript and a 1.4-kb DuraScript RNA transcript were each incubated with 1 U of highly purified RNase A for 30 minutes. The standard RNA transcript was completely degraded while the DuraScript RNA transcript remained intact. Lane M, size ladder; lane 1, 1.4-kb standard RNA transcript; lane 2, standard RNA after RNase A treatment; lane 3, 1.4-kb DuraScript RNA; lane 4, DuraScript RNA after RNase A treatment.
Figure 4. DuraScript® RNA is completely resistant to “finger” nucleases. A 1.4-kb standard RNA transcript and a 1.4-kb DuraScript RNA transcript were produced using sterile water or water that had been contaminated by exposure to the hands of a test subject. The standard RNA shows extensive degradation from “finger” nucleases in the contaminated water while the DuraScript RNA remains fully intact. M, Size ladder; Lane 1, Standard RNA transcript; Lane 2, Standard RNA after “finger” nuclease exposure; Lane 3, DuraScript RNA; Lane 4, DuraScript RNA after “finger” nuclease exposure.
* Covered by issued and/or pending patents.
Ordering
Catalog No. | Size |
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DuraScribe® T7 Transcription Kit
DS010910 | 10 Reactions |
DS010925 | 25 Reactions |