Hi Mike, For long RNAs (> 40bases), in vitro transcription is the method of choice. You might want to take a look at this introductory page from Ambion:
http://www.ambion.com/techlib/basics/transcription/index.html For structural studies you will have to scale up to milliliter scale. For that you might want to produce your own T7 RNA polymerase. Another thing to consider is the method of purification of your RNA. I believe purification from denaturing gel is the most widely used method, but you should consider native chromatographic purification as well. The following references should get you started on the subject: http://www.ncbi.nlm.nih.gov/pubmed/20946782 http://www.ncbi.nlm.nih.gov/pubmed/17272845 If you absolutely want to go with chemical synthesis, I believe you will have to buy shorter RNA pieces (40 bases or less) and ligate them later using a T4 RNA ligase. There are many good references out there that deal with production, purification and crystallization or RNA. Good luck, Mario Sanches On Sun, Mar 13, 2011 at 4:39 AM, Michael Thompson <[email protected]>wrote: > Hello All, > > I am looking for some advice from some experienced RNA crystallographers. I > would like to order some relatively short (<90 bases) synthetic RNAs for > crystallization trials. I was wondering if anyone could comment on the use > of synthetic RNAs for crystallization. Specifically, what is the longest > synthetic RNA that can be used for crystallization trials? I've seen some > structures in the PDB that are up to 88 bases and are reported to have been > obtained with synthetic constructs (3OWI - glycine riboswitch), but I don't > really know if that's routine or if it's an exceptional case. Also, for > those who have experience with the use of synthetic RNAs, I was wondering > where people generally order their synthetic constructs from? Our resident > expert in RNA crystallography recommended a company called Dharmacon (part > of ThermoFisher), but I was hoping that I might get some other opinions as > to which companies make the best quality oligonucleotides, provide samples > with the highest purity, and have the most reasonable prices. > > Thanks in advance for the help! > > Mike > > > > -- > Michael C. Thompson > > Graduate Student > > Biochemistry & Molecular Biology Division > > Department of Chemistry & Biochemistry > > University of California, Los Angeles > > [email protected] > -- Mario Sanches Postdoctoral Fellow Samuel Lunenfeld Research Institute Mount Sinai Hospital 600 University Ave Toronto - Ontario Canada M5G 1X5 http://ca.linkedin.com/in/mariosanches
