Nucleotides for Applications on RNA

in vitro Transcription Grade Ultrapure Nucleoside Triphosphates

In vitro synthesis of RNA is catalyzed by bacteriophage (SP6, T7 or T3) RNA polymerases using linear DNA as a template. Ribonucleoside triphosphates (NTPs) are the building blocks of these in vitro transcription → Molecular Biology reactions.

Since amplification sensitivity, product yield and reproducibility are strongly affected by contaminating impurities, the quality of NTPs is of considerably importance.

Each lot of Jena Bioscience’s ultrapure (> 99 %) NTPs is confirmed free of DNase, RNase, Protease and nicking activity. Furthermore, all NTPs are functionally tested by T7 RNA Polymerase-mediated in vitro transcription.

Name Cat. No. Size
ATP – Solution NU-1010 1 ml (100 mM)
CTP – Solution NU-1011 1 ml (100 mM)
GTP – Solution NU-1012 1 ml (100 mM)
UTP – Solution NU-1013 1 ml (100 mM)

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Unmodified (natural) ribo-Nucleoside triphosphates (NTPs) – Bundles

In vitro synthesis of RNA is catalyzed by bacteriophage (SP6, T7 or T3) RNA polymerases using linear DNA as a template. Ribonucleoside triphosphates (NTPs) are the building blocks of these in vitro transcription → Molecular Biology reactions.

Since amplification sensitivity, product yield and reproducibility are strongly affected by contaminating impurities, the quality of NTPs is of considerably importance.

Each lot of Jena Bioscience’s ultrapure (> 99 %) NTPs is confirmed free of DNase, RNase, Protease and nicking activity. Furthermore, all NTPs are functionally tested by T7 RNA Polymerase-mediated in vitro transcription.

Name Cat. No. Size
NTP Bundle NU-1014S 4 x 200 μl (4 x 20 μmol)
NTP Bundle NU-1014L 4 x 1 ml (4 x 100 μmol)

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Unmodified (natural) ribo-Nucleoside triphosphates (NTPs) – Solids

In vitro synthesis of RNA is catalyzed by bacteriophage (SP6, T7 or T3) RNA polymerases using linear DNA as a template. Ribonucleoside triphosphates (NTPs) are the building blocks of these in vitro transcription → Molecular Biology reactions.

Since amplification sensitivity, product yield and reproducibility are strongly affected by contaminating impurities, the quality of NTPs is of considerably importance.

Each lot of Jena Bioscience’s NTP powders is confirmed free of DNase, RNase, Protease and nicking activity. Furthermore, all NTPs are functionally tested by T7 RNA Polymerase-mediated in vitro transcription.

Name Cat. No. Size
ATP – Solid NU-1010-1G 1 g
ATP – Solid NU-1010-10G 10 g
ATP – Solid NU-1010-100G 100 g
CTP – Solid NU-1011-100 100 mg
CTP – Solid NU-1011-1G 1 g
CTP – Solid NU-1011-10G 10 g
CTP – Solid NU-1011-100G 100 g
GTP – Solid NU-1012-100 100 mg
GTP – Solid NU-1012-1G 1 g
GTP – Solid NU-1012-10G 10 g
GTP – Solid NU-1012-100G 100 g
GTP – Solid – Purity 85 % NU-1047-200 200 mg
GTP – Solid – Purity 85 % NU-1047-1G 1 g
GTP – Solid – Purity 85 % NU-1047-10G 10 g
GTP – Solid – Purity 85 % NU-1047-100G 100 g
UTP – Solid NU-1013-100 100 mg
UTP – Solid NU-1013-1G 1 g
UTP – Solid NU-1013-10G 10 g
UTP – Solid NU-1013-100G 100 g

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Cap Analogs – Enhance mRNA stability and translation efficiency

Many eukaryotic and viral mRNAs are modified at their 5′ ends by addition of 7-Methylguanosine (N7-methyl guanosine or m7G), known as “Cap”. “Capping” of the mRNA structure plays a crucial role in a variety of cellular processes which include translation initiation[1], splicing[2], intracellular transport[3] and turnover[4].

Consistently, successful downstream application of in vitro transcribed mRNAs strongly depends on the 5′ Cap structure. Capped mRNAs are generally more efficiently translated in wheat germ and reticulocyte in vitro translation systems[5], and they are less susceptible to exonuclease degradation during microinjection experiments compared to uncapped mRNAs[6].

In vitro synthesis of capped mRNAs is performed by bacteriophage RNA polymerase (T7, SP6 or T3)-mediated in vitro transcription that co-transcriptionally incorporate cap analogs at the 5′-end of the transcripts (Fig. 1).

Reactions with traditional cap analogs (GpppG, m7GpppG or m2,2,7GpppG) routinely yield a mixture of mRNAs containing the cap analog incorporated both in a correct or reverse orientation[7]. Thus about 50 % of the resulting capped mRNAs are recognized by the translational machinery. Translational efficiency however, can be markedly increased by usage of the “anti-reverse” cap analog (ARCA, m7,3′-OGpppG)[8]. This is due to substitution of the 3′-OH of the m7 guanine moiety by a 3′-O-methyl group that forces ARCA incorporation in the correct orientation and subsequently results in a 100 % translatable mRNA population.

Figure 1: Cap analogs are enzymatically incorporated at the RNA 5′-end by bacteriophage RNA Polymerase-mediated in vitro Transcription.

Name Cat. No. Size
GP3G – Solid (Unmethylated Cap Analog) NU-854-1 1 mg
GP3G – Solid (Unmethylated Cap Analog) NU-854-5 5 mg
GP3G (Unmethylated Cap Analog) – Solution NU-854S 10 μl (100 mM)
GP3G (Unmethylated Cap Analog) – Solution NU-854L 5 x 10 μl (100 mM)
m7GP3G (Monomethylated Cap Analog) NU-852-1 1 mg
m7GP3G (Monomethylated Cap Analog) NU-852-5 5 mg
m7GP3G (Monomethylated Cap Analog) – Solution NU-852S 10 μl (100 mM)
m7GP3G (Monomethylated Cap Analog) – Solution NU-852L 5 x 10 μl (100 mM)
m32.2.7GP3G (Trimethylated Cap Analog) NU-853-1 1 mg
m32.2.7GP3G (Trimethylated Cap Analog) NU-853-5 5 mg
m27,3′-OGP3G (ARCA Cap Analog) – Solid NU-855-1 1 mg
m27,3′-OGP3G (ARCA Cap Analog) – Solid NU-855-1 1 mg
m27,3′-OGP3G (ARCA Cap Analog) – Solid NU-855-5 5 mg
m27,3′-OGP3G (ARCA Cap Analog) – Solution NU-855S 10 μl (100 mM)
m27,3′-OGP3G (ARCA Cap Analog) – Solution NU-855L 5 x 10 μl (100 mM)

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Selected References

[1] Gingras et al. (1999) eIF4 initiation factors: Effectors of mRNA recruitment to ribosomes and regulators of translation. Annu. Rev. Biochem. 68:913.
[2] Izaurralde et al. (1994) A nuclear cap binding protein complex involved in pre-mRNA splicing. Cell 78:657.
[3] Izaurralde et al. (1992) A cap binding protein that may mediate nuclear export of RNA polymerase II-transcribed RNAs. J. Cell Biol. 118:1287.
[4] Beelman et al. (1998) An essential component of the decapping enzyme required for normal rates of mRNA turnover. Nature 382:642.
[5] Paterson et al. (1979) Efficient translation of prokaryotic mRNAs in a eukaryotic cell-free system requires addition of a cap structure. Nature 279:692.
[6] Drummond et al. (1985) The effect of capping and polyadenylation on the stability, movement and translation of synthetic messenger RNAs in Xenopus oocytes. Nucl. Acids Res. 13:375.
[7] Pasquinelli et al. (1995) Reverse 5′ caps in RNAs made in vitro by phage RNA polymerases. RNA 1:957.
[8] Grudzien et al. (2007) Synthesis of Anti-Reverse Cap Analogs (ARCAs) and their Applications in mRNA Translation and Stability. Methods Enzymol. 431:203.

Nucleotides for Structure Determination by NMR

RNA plays a fundamental role in biological processes (e.g. translation or gene regulation). Therefore, the biophysical properties of RNA molecules such as their three dimensional shape or secondary structure distribution gained increasing importance.

RNA structure determination can be performed by liquid state nuclear magnetic resonance (NMR) spectroscopy that requires millimolar amounts of isotopically labeled RNA to obtain well-resolved signals.

A successful approach to generate sufficient isotopically labeled RNA amounts is the in vitro Transcription-mediated synthesis of 19F-labeled RNA using fluorinated NTPs[1]. 2F-ATP or 5F-UTP are incorporated instead of their natural counterparts in a non-perturbing way (intact base-pairing properties) and simultaneously function as a sensitive NMR reporter group (larger chemical shift dispersion than 1H)[1].

 

Name Cat. No. Size
2-Fluoro-ATP NU-145S 10 μl (100 mM)
2-Fluoro-ATP NU-145L 5 x 10 μl (100 mM)
5-Fluoro-UTP NU-1115S 10 μl (100 mM)
5-Fluoro-UTP NU-1115L 5 x 10 μl (100 mM)

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Selected References

[1] Roth et al. (2011) RNA Structure Determination by NMR: Combining Labeling and Pulse Techniques. Advances in Biomedical Spectroscopy 3:205.

Nucleotides for RNA Crosslinking

Nucleotides for RNA functionalization and subsequent crosslinking.

For application references, please refer to the corresponding datatsheet.

Name Cat. No. Size
4-Thio-UTP NU-1156S 10 μl (100 mM)
4-Thio-UTP NU-1156L 5 x 10 μl (100 mM)
5-Bromo-UTP NU-121S 50 μl (10 mM)
5-Bromo-UTP NU-121L 5 x 50 μl (10 mM)
5-Azido-C3-UTP NU-157S 20 μl (10 mM)
5-Azido-C3-UTP NU-157L 5 x 20 μl (10 mM)
BzBzATP (BzATP) NU-1620-5 5 mg
BzBzATP (BzATP) NU-1620-25 25 mg
8-Azido-ATP NU-155S 500 μl (10 mM)
8-Azido-ATP NU-155L 5 x 500 μl (10 mM)
γ-(6-Azidohexyl)-ATP NU-1702S 50 μl (10 mM)
γ-(6-Azidohexyl)-ATP NU-1702L 5 x 50 μl (10 mM)
γ-(2-Azidoethyl)-ATP NU-1701S 100 μl (10 mM)
γ-(2-Azidoethyl)-ATP NU-1701L 5 x 100 μl (10 mM)

Bulk Amounts

If you require large amounts (> 10 ml, > 100 ml, > 1 l), significant discounts are available. Please contact us at info@stratech.co.uk to receive an individual quotation.

Selected References

Luo et al. (2003) Identification of RNA Binding Proteins by UV Cross-Linking. Current Protocols in Molecular Biology 27.2.1.

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