Navigation auf uzh.ch

Suche

Department of Biochemistry

Microseeding

Two detailed protocols on how to prepare a microseed stock can be found here:

“Procedure for making the microseed stock” from Patrick D. Shaw Stewart (Douglas Instruments Ltd).   
Print friendly version (PDF) (PDF, 1 MB)

“Matrix Seeding” from Janet Newman (Collaborative Crystallisation Centre (C3)).

The preparation of the microseed stock will be carried out by the researcher. PCC will assist in the preparation of the microseed stock by providing “Seed Beads”(Hampton Research, HR2-320) and a “Micro-Tools Set”.
The initial steps for setting up a microseeding experiment is identical to the vapor diffusion crystallization experiment described in the section Experimental Setup.
The addition of the microseed stock to each drop of the vapor diffusion crystallization experiment is carried out by the Gryphon-LCP at 20°C or 4°C. The researcher specifies the volume of the microseed stock to be added to the three crystallization wells. The microseed stock volume added is commonly in the range between 20nl and 40nl.
The volume for the microseed stock required for setting up a crystallization plate can be found in Sample Volume.

The Oxford Protein Production Facility-UK provides an Automated Microseed Protocol for Protein Crystallization

The following list of microseeding papers is an introduction to the microseeding crystallization technique:

list of papers

Allan D’Arcy, et al.
Microseed matrix screening for optimization in protein crystallization: what have we learned?; Acta Crystallogr F Struct Biol Commun. 2014 Sep;70(Pt 9):1117-26.

Allan D’Arcy, et al.
An automated microseed matrix-screening method for protein crystallization;
Acta Crystallogr D Biol Crystallogr. 2007 Apr;63(Pt 4):550-4.

Marisa Till, et al.
Improving the success rate of protein crystallization by random microseed matrix screening.n; J Vis Exp. 2013 Aug 31;(78).

Gregory C.Ireton, et al.
Microseed matrix screening to improve crystals of yeast cytosine deaminase;
Acta Cryst. (2004). D60, 601-605

Galina Obmolova, et al.
Promoting crystallization of antibody–antigen complexes via microseed matrix screening; Acta Cryst. (2010). D66, 927–933

Armando G. Villasenor, et al.
Acoustic matrix microseeding: improving protein crystal growth with minimal chemical bias; Acta Cryst. (2010). D66, 568–576

Patrick D. Shaw Stewart, et al.
Random Microseeding: A Theoretical and Practical Exploration of Seed Stability and Seeding Techniques for Successful Protein Crystallization;
Cryst. Growth Des. 2011, 11, 3432–3441

Abuhammad Areej, et al.
Structure of arylamine N-acetyltransferase from Mycobacterium tuberculosis ..."; Acta Crystallographica Section D: Biological Crystallography 69, no. 8 (2013): 1433-1446

Enrico A. Stura, et al.
Epitaxial jumps; Journal of crystal growth 196, no. 2 (1999): 250-260.

Stefan A. Kolek, et al.
A novel microseeding method for the crystallization of membrane proteins in lipidic cubic phase;
Acta Crystallographica Section F, Volume 72; Part 4; April 2016; Pages 307-312.

see also: RAMC2011_T11_Obmolova (PDF, 1 MB)

Weiterführende Informationen

PCC-Pinboard

New employee at PCC

I am delighted to welcome Görkem Kurtuldu to PCC as a Senior Scientist. She will be stepping into the role as my designated successor. Görkem brings a fresh perspective to PCC, and I am excited to see the positive impact she will make.
Please join me in welcoming Görkem!

New screen:

What’s new:
PCC supports researchers with preparing custom screens through their pipetting service. The researcher can freely compose a screen with up to 30 different stock solutions.
Be sure to have a look and feel free to ask the PCC staff concerning this new service.

We had to adjust the prices for plates that are set up at 4°C due to the significantly higher wear and tear of the liquid handler stationed in the cold room.

The new screen Morpheus® Fusion (Product#: MD1-129) from Molecular Dimensions is now available at PCC as the initial screen GS087.

 

RockImager and Liquid Handler Status

RockImager 20°C:
The imaging schedule is on time.

RockImager 4°C:
The imaging schedule is on time.

Crystal Gryphon 4°C:
In Operation.

Crystal Gryphon LCP 20°C:
In Operation.

Crystal Phoenix 20°C:
In Operation.

Freedom EVO w/ MCA 96 + RoMa
In Operation

Freedom EVO w/ LiHa
In Operation

Contact

Department of Biochemistry
University of Zurich
Winterthurerstr. 190
CH-8057 Zurich
Office: Y44 J 30/34
xtal@bioc.uzh.ch


Görkem Kurtuldu
Tel.: +41 44 635 6557