Hi Saurabh Of course you should use "rMMS" microseeding. See below
I hope it works ! Best wishes, Patrick ---------- Forwarded message --------- From: Patrick Shaw Stewart <patr...@douglas.co.uk> Date: Mon, Sep 9, 2019 at 10:12 AM Subject: Re: [ccp4bb] Optimization from needle shaped crystals To: David Briggs <david.bri...@crick.ac.uk> Cc: CCP4BB@JISCMAIL.AC.UK <CCP4BB@jiscmail.ac.uk> Hi Chitra Needles usually work very well for making seed stocks for random Microseed Matrix Screening (MMS). Your protein probably crystallizes well, but it is growing too fast in one direction. MMS has lots of advantages. If it's going to work it will almost certainly work within 12 hours. Also, it allows you to control the number of crystals per drop by diluting the seed stock. Another excellent (open-access) paper that gives lots of examples of seeding, cross-seeding, dilution, optimization etc within a defined set of fabs is here: https://scripts.iucr.org/cgi-bin/paper?nj5193 Good luck, Patrick On Sun, Sep 8, 2019 at 1:46 PM David Briggs <david.bri...@crick.ac.uk> wrote: > 4. Matrix microseeding. Make a seed stock from these crystals and then > re-run your primary screens. > > https://www.ncbi.nlm.nih.gov/m/pubmed/25195878/ > > -- > Dr David C. Briggs > Senior Laboratory Research Scientist > Signalling and Structural Biology Lab > The Francis Crick Institute > London, UK > == > about.me/david_briggs > > On Sun, Mar 21, 2021 at 6:04 PM Saurabh Upadhyay <saurabh807...@gmail.com> wrote: > Dear All, > > I am trying to crystalize a protein, which occurs in monomeric (60kDa) and > tetrameric (240kDa) form. *The protein during crystallization was in > MES buffer pH-6 *and the method of crystallization was "*Sitting drop*" > in "*Proplex*" condition. Some of the conditions in which crystals were > obtained are: > > *1) 0.1M HEPES pH-7; 10% (w/v) PEG 4000* > *2) 0.1M Tris, pH-8; 8% (w/v) PEG 8000* > *3) 0.1M Tris, pH-8; 25% (v/v) PEG 400* > > For reference, I have attached the images of the crystals observed below. > > *However, the diffraction pattern obtained and the resolution of crystals > were very poor. Even some mosaicity has been observed. * > > Kindly suggest some methods or modifications, to improve the resolution > and diffraction pattern of the crystals obtained. > > Thanking You, > Sincerely, > Saurabh Upadhyay, > Ph.D. Scholar > c/o Dr. Ashok kumar Patel, > Kusuma School of Biological Sciences, > Indian Institute of Technology, Hauz Khas, > New Delhi-110016, India > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > -- patr...@douglas.co.uk Douglas Instruments Ltd. Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK Directors: Patrick Shaw Stewart, Peter Baldock, Stefan Kolek http://www.douglas.co.uk Tel: 44 (0) 148-864-9090 US toll-free 1-877-225-2034 Regd. England 2177994, VAT Reg. GB 480 7371 36 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
