Hi folks just my two ha’porth.
Back in the mid 1990s, when MAD was becoming common and tunable beamlines were being installed at every synchrotron you could shake a stick at, I was involved in several successful projects involving 5-Br-U in oligo-DNA crystallography. In my (very, very) naïve hands at the time it worked like magic (I remember my shock at seeing the base stacking in the first map I calculated for one of the structures) - and this was using data collected on image plates with each image spanning rather more than 1 degree - so not how it would be done now. Since just about everything to do with data collection and processing and structure solution has improved by leaps and bounds since then, I would back Br-SAD to yield a structure. If anyone’s interested, they can see my contribution regarding this at the 1997 CCP4 Study Weekend proceedings (this was prior to them appearing in Acta D), available at - https://legacy.ccp4.ac.uk/courses/proceedings/1997/h_powell/main.html Contributions at the same meeting by others whose names will be instantly recognizable may well be more use than mine… Best wishes Harry > On 19 Sep 2023, at 19:32, Wagner, Armin (DLSLtd,RAL,LSCI) > <0000ada38eff8538-dmarc-requ...@jiscmail.ac.uk> wrote: > > 5-Br-U is a good alternative and works well, but we have also managed to > solve RNA/DNA by K-SAD or Co-SAD (https://doi.org/10.1093/nar/gkaa439) and Ca > could be attractive as a potential anomalous scatter as well. > > Best regards, > > Armin > > > > > From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Mark J. van > Raaij <mjvanra...@cnb.csic.es> > Date: Tuesday, 19 September 2023 at 12:31 > To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> > Subject: Re: [ccp4bb] the structures of Nucleic acid > > This just appeared and may be relevant: > https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkad726/7272628 > > Critical Reviews and Perspectives > When will RNA get its AlphaFold moment? > Mark van Raaij > Dpto de Estructura de Macromoleculas, lab 20B > Centro Nacional de Biotecnologia - CSIC > calle Darwin 3 > E-28049 Madrid, Spain > tel. +34 91 585 4616 (internal 432092) > > > > On 18 Sep 2023, at 18:07, William G. Scott > <00002844d921eb97-dmarc-requ...@jiscmail.ac.uk> wrote: > > The phosphorus absorption edge is about 5.8Å. > > I've had much better luck with 5-Br-U for anomalous phasing. > > Molecular replacement with sub-structural fragments can also work: > <https://scottlab.ucsc.edu/scottlab/reprints/2010_Scott_Methods.pdf> > > > Yours sincerely, > > William G. Scott > Professor, Department of Chemistry and Biochemistry > and The Center for the Molecular Biology of RNA > University of California at Santa Cruz > Santa Cruz, California 95064 > USA > > > On Sep 18, 2023, at 2:43 AM, Eleanor Dodson > <0000176a9d5ebad7-dmarc-requ...@jiscmail.ac.uk> wrote: > > I am afraid most scientists will use the most straightforward technique! > If SAD is available the PHOSPHATE backbone of DNA will provide sufficient > signal to allow SAD to work, and you get an unambiguous answer to whether it > is A-DNA or B or Z... > MR will usually work of course as well > Eleanor > > > On Mon, 18 Sept 2023 at 09:18, Natesh Ramanathan <nat...@iisertvm.ac.in> > wrote: > Dear Fu Xingke, > > Depends on what Nucleic Acid you are talking of. If it is RNA, you > can expect some sequence to tertiary structure correspondence so you might be > able to try more MR as compared to DNA. DNA may have double helical > architecture but less sequence to tertiary structure correspondence, and > hence DNA is less likely to have a 3D structure like RNA specific structure > for a sequence. > > SAD has become a straight forward method to avoid all these problems > to get ab-initio structure. So many go for it directly. > > Hope that helps. > Best wishes, > Natesh > > On Mon, 18 Sept 2023 at 13:36, fuxingke <fuxingke0...@163.com> wrote: > Dear Colleagues, > > Reacently, I find the structures of Nucleic acid are solved by > single-wavelength anomalous diffraction(SAD). So, why molecular replacement > (MR) not? > > Regards > > > > Best wishes, > > Fu Xingke > > Institute of Physics CAS > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > > > > -- > ---------------------------------------------------------- > "Live Simply and do Serious Things .. 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