Stable Isotope Labeled Nucleotide Triphosphates & DNA Phosphoramidites
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1 Cambridge Isotope Laboratories, Inc. Cambridge Isotope Laboratories, Inc. RESEARCH PRODUCTS Stable Isotope Labeled Nucleotide Triphosphates & DNA Phosphoramidites NMR spectroscopy is an extremely powerful and versatile tool for studying the dynamics and structure of RNA and DNA molecules in solution. The types of information gained from NMR studies of RNA and DNA include 1 : -pairing pattern Conformational equilibria Site-specific information regarding ligand binding Delineation of secondary structure motifs, such as hairpins and bulges Local structure and dynamics Global structure derived from rdcs
2 Cassia LLC was founded in 2005 by Dr. Lincoln Scott and noted NMR spectroscopist Dr. Jamie Williamson. CIL and Cassia have a special relationship which makes use of CIL s isotopic material production and marketing and Cassia s special knowledge of RNA and DNA biosynthesis. Since 2005, CIL and Cassia have developed the most extensive product line of stable isotope labeled RNA and DNA triphosphates, DNA phosphoramidites and other related compounds. All of these products are routinely available for immediate shipment from stock at CIL, Inc. For more information on any labeled RNA or DNA products contact your local CIL representative. We have enjoyed a close working relationship with CIL for the last 15 years, both as a customer and a collaborator. We ve had great interactions with Sales, Management and the Chemists from top to bottom. CIL is a great company that you can really work with in this specialized area. We ve been able to do science that we couldn t have done without working with CIL. James R. Williamson, Ph.D. Professor Dean of Graduate Studies Departments of Molecular Biology and Chemistry The Skaggs Institute for Chemical Biology The Scripps Research Institute CIL is proud to offer RNA and DNA products manufactured by or in conjunction with Cassia, LLC
3 Cambridge Isotope Laboratories, Inc. RNA TRIPHOSPHATES The most popular approaches to produce labeled RNA molecules for NMR studies use enzymatic in vitro transcription methods that employ labeled rntps, T7-RNA polymerase and either linearized plasmids or double-stranded DNA as templates. These techniques are used to construct labeled RNA molecules of which all of one type of nucleotide is labeled. RNA TRIPHOSPHATES UNIFORMLY D LABELED Perdeuterated NTPs can be used in combination with protonated NTPs to create RNA molecules in which specific types of nucleotides are protonated, thus allowing spectral editing without the significant signal broadening associated with 13 C incorporation 2. DLM-7514-CA Adenosine 5 -Triphosphate, Ammonium Salt (U-D, 97%+) DLM-7515-CA Cytidine 5 -Triphosphate, Ammonium Salt (D 8, 97%+) DLM-7516-CA Guanosine 5 -Triphosphate, Ammonium Salt (U-D, 97%+) DLM-7517-CA Uridine 5 -Triphosphate, Ammonium Salt (D 8, 97%+) DLM-7518-CA Set of four Ribonucleoside -5 -Triphosphates, Ammonium Salt (U-D, 97%+) Note: All products are in solution and have a chemical purity >90% SELECTIVELY DEUTERATED Because severe signal degeneracy has hampered NMR studies of larger RNAs, key researchers in this area have utilized selectively deuterated rntps, in conjunction with in vitro synthesis methods, to reduce spectral complexity, spectral line-widths, and for observing NOEs over larger distances 2. DLM-7862 Equimolar Mix:ATP, GTP (ribose-3,4,5,5 -D 4,98%); CTP, UTP (5-D 1, ribose-3,4,5,5 - D 4,98%) DLM-8594 Cytidine 5 Triphosphate (cytosine-5-d, 6-H; ribose-1,2,3,4,5,5-d 6, 96-97%) DLM-8637 Uridine 5 Triphosphate (uracil-5-d, 6-H; ribose-1,2,3,4,5,5-d 6, 96-97%) UNIFORMLY 15 N LABELED NLM-3987-CA Adenosine 5 -Triphosphate, Ammonium Salt (U- 15 N 5 NLM-4266-CA Cytidine 5 -Triphosphate, Ammonium Salt (U- 15 N 3 NLM-4268-CA Guanosine 5 -Triphosphate, Ammonium Salt (U- 15 N 5 NLM-4270-CA Uridine 5 -Triphosphate, Ammonium Salt (U- 15 N 2 NLM-8772-CA Set of four Ribonucleotide 5 -Triphosphates, Ammonium Salt (U- 15 N Note: All products are in solution and have a chemical purity >90% UNIFORMLY 13 C LABELED ( 13 C, 98% Enrichment) CLM-8426 Adenosine 5 -Triphosphate CLM-8427 Cytidine 5 -Triphosphate CLM-8428 Guanosine 5 -Triphosphate CLM-8429 Uridine 5 -Triphosphate CLM x50 Set of four Ribonucleotide 5 -Triphosphates (U- 13 C, 98%) UNIFORMLY 13 C/ 15 N LABELED CNLM-4265-CA Adenosine 5 -Triphosphate, Ammonium Salt (U- 13 C, U- 15 N CNLM-4267-CA Cytidine 5 -Triphosphate, Ammonium Salt (U- 13 C, U- 15 N CNLM-4269-CA Guanosine 5 -Triphosphate, Ammonium Salt (U- 13 C, U- 15 N CNLM-4271-CA Uridine 5 -Triphosphate, Ammonium Salt (U- 13 C, U- 15 N CNLM-7503-CA Set of four Ribonucleoside 5 -Triphosphates, Ammonium Salt (U- 13 C, U- 15 N Note: All products are in solution and have a chemical purity >90%
4 DNA TRIPHOSPHATES Labeled DNA oligonucleotides are routinely synthesized using enzymatic in vitro methods that require labeled dntps, a DNA polymerase, and a cdna template. One particular advantage of using enzymatic methods over synthetic chemistry methods is that large oligonucleotides (e.g., >50 nucleotides in length) can be easily prepared in milligram quantities. DNA TRIPHOSPHATES UNIFORMLY D LABELED (D, 97% + Enrichment) DLM Deoxyadenosine 5 -Triphosphate DLM Deoxycytidine 5 -Triphosphate DLM Deoxyguanosine 5 - Triphosphate DLM-7510 Thymidine 5 -Triphosphate DLM-7511 Set of four Deoxyribonucleoside 5 -Triphosophates Uniformly 15 N Labeled ( 15 N, 96-99% Enrichment) NLM Deoxyadenonsine 5 -Triphosphate NLM Deoxycytidine 5 -Triphosphate NLM Deoxyguanosine 5 -Triphosphate NLM-6218 Thymidine 5 -Triphosphate NLM-7512 Set of four 2 -Deoxyribonucleoside-5 -Triphosphate Uniformly 13 C Labeled ( 13 C, 98% Enrichment) CLM Deoxyadenosine 5 -Triphosphate CLM Deoxycytidine 5 -Triphosphate CLM Deoxyguanosine-5 -Triphosphate CLM-8433 Thymidine-5 -Triphosphate CLM x50 Set of four 2 -Deoxyribonucleotide 5 -Triphosphates (U- 13 C, 98%) Uniformly 13 C/ 15 N Labeled ( 13 C, 98%; 15 N, 98%) CNLM Deoxyadenosine 5 -Triphosphate CNLM Deoxycytidine 5 -Triphosphate CNLM Deoxyguanosine 5 -Triphosphate CNLM-6222 Thymidine 5 -Triphosphate CNLM-7513 Set of four 2 -Deoxyribonucleoside 5 -Triphosphate
5 Cambridge Isotope Laboratories, Inc. DNA PHOSPHORAMIDITES Position-specific labeled DNA molecules can be synthesized using standard phosphoramidite chemistry to overcome the limited chemical-shift dispersion of DNA, as well as to obtain residue-specific functional, structural and dynamic information. DNA PHOSPHORAMIDITES Uniformly 15 N Labeled ( 15 N,98%) NLM Deoxyadenosine Phosphoramidite NLM Deoxycytidine Phosphoramidite NLM Deoxyguanosine Phosphoramidite NLM-6823 Thymidine Phosphoramidite Uniformly 13 C/ 15 N Labeled ( 13 C, 98%; 15 N, 98%) CNLM Deoxyadenosine Phosphoramidite CNLM Deoxycytidine Phosphoramidite CNLM Deoxyguanosine Phosphoramidite CNLM-6824 Thymidine Phosphoramidite CNLM-6824 Thymidine Phosphoramidite CNLM Deoxycytidine Phosphoramidite CNLM Deoxyadenosine Phosphoramidite CNLM Deoxyguanosine Phosphoramidite 1 Furtig, B; Richter, C; Wohnert, J; Schwalbe, H. NMR Spectroscopy of RNA (2003) ChemBioChem 4: Lu, K; Miyazaki, Y; Summers, M. Isotope Labeling Strategies for NMR Studies of RNA (2010) J Biomol NMR 46:
6 Top 10 Reasons to Use Ammonium Salts Self-Buffering (ph ~ 7.6) Soft Cation Nucleotides of ammonium salts are active with polymerases, synthetases, and phosphatases. Volatile counter-ion The ammonium cation can be easily exchanged using DOWEX cation exchange resin The ph does not change during drying of the nucleotide (i.e. speed -vac, lyophilize) Stoichiometry between the counter-ion and the nucleotide is preserved Routinely compatible in down-stream syntheses Compatible in a variety of down-stream chromatography applications Tested to be comparable in side-by-side in vitro transcription reactions! To request a quotation or place an order, please contact CIL Sales at cilsales@isotope.com telephone: (N.America only) For our international customers please contact CIL International Sales at intsales@isotope.com telephone: CIL s distributor listing is available at: isotope.com Cambridge Isotope Laboratories, Inc. 50 Frontage Road Andover, MA RNA2010
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