Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden...
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![Page 1: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/1.jpg)
Seeding
Terese Bergfors
Dept of Cell and Molecular Biology
Uppsala University, Uppsala Sweden
Examples of two new structures from our
laboratory where seeding was critical to obtaining the
crystals
![Page 2: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/2.jpg)
Seeding
• Why seed?
• How?
• What to use as a seed?
![Page 3: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/3.jpg)
Some methods for optimization
microgravity seeding
![Page 4: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/4.jpg)
Seeding and the phase diagram
Luft and De Titta, Acta Cryst. (1999) D55, 988-993
![Page 5: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/5.jpg)
Serial seeding
20 mg/ml
10 mg/ml
10 mg/ml
![Page 6: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/6.jpg)
What to seed from?
oil
Take your best crystal
or
any solid phase
![Page 7: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/7.jpg)
Streak seedingwhisker
Touchcrystal
Draw streak line
Enrico Stura in “Crystallization of Nucleic Acids and Proteins” Ed. Ducruix and Giege,
1992 Oxford University Press p. 108
![Page 8: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/8.jpg)
Does seeding solve everything?
before after
![Page 9: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/9.jpg)
Does seeding solve everything?
After seeding, Ni, and ligand
• After Seeding, Ni
![Page 10: Seeding Terese Bergfors Dept of Cell and Molecular Biology Uppsala University, Uppsala Sweden terese@alpha2.bmc.uu.se Examples of two new structures from.](https://reader033.fdocuments.in/reader033/viewer/2022052509/56649d2c5503460f94a017e1/html5/thumbnails/10.jpg)
Summary of Tips
• Lower concentration
• Use any solid phase
• Serial seed
• Use sitting drops