Seeding allows one to grow crystals in the metastable zone. Crystallization in this zone provides control, reproducibility and an improved likelihood of a successful crystallization experiment. Also, crystals can grow from seeds but cannot spontaneously nucleate. By placing a seed or solution of seeds in a drop which is saturated to the metastable zone, one can use the seeds to grow larger single crystals. By controlling the number of seeds introduced into the drop, one can control the number of crystals grown. It is not practically possible to measure and know the number of seeds introduced to a drop, but by performing serial dilutions from a concentrated seed stock, one can control the number of crystals grown in the drop.

Using the Seed Bead kits, one can create crystal seed stock for subsequent seeding experiments. Crystals are placed in the microcentrifuge tube with the Seed Bead(s) and either vortexed or sonicated to generate a seed stock. Then by performing serial dilutions, one can control the number and size of crystals in the experiment.

The Seed Bead kit is useful for the preparation of seed stocks for automated and semi-automated microseeding (D’Arcy 2007, Harlos 2008).

Each Seed Bead Kit (HR2-320) contains 24 special microcentrifuge tubes, each tube containing a single 3mm PTFE Seed Bead.

Each Seed Bead Steel Kit (HR4-780) contains 24 special microcentrifuge tubes, each tube containing a six 1.6 mm Stainless Steel Seed Beads.

Each Seed Bead Ceramic Kit (HR4-781) contains 24 special microcentrifuge tubes, each tube containing a ten 1.0 mm Zirconium Silicate Ceramic Seed Beads.

Each Seed Bead Glass Kit (HR4-782) contains 24 special microcentrifuge tubes, each tube containing a ten 1.0 mm Glass Seed Beads.

Seed Bead™ 试剂盒
应用
产生蛋白质晶体的种子
特征
轻松生成一致的种子库存
使用连续稀释来控制引入液滴的种子数量
微种子基质筛选 (MMS) 种子储备
PTFE、不锈钢、硅酸锆陶瓷或玻璃种子珠
描述
Seed Bead 试剂盒用于创建种子库以进行后续的播种实验。 Seed Bead 试剂盒包含 24 个 Seed Bead Tubes，这是一种特殊的 1.5 ml 微量离心管，其形状可在混合过程中将种子保持在样品中。每个管都配置有 PTFE、不锈钢、硅酸锆陶瓷或玻璃种子珠。

接种允许人们在亚稳态区域中生长晶体。该区域中的结晶提供了控制、可重复性和成功结晶实验的改进可能性。此外，晶体可以从种子生长，但不能自发成核。通过将种子或种子溶液放入饱和到亚稳区的液滴中，可以使用种子生长更大的单晶。通过控制引入液滴的种子数量，可以控制晶体生长的数量。实际上不可能测量和知道引入一滴的种子数量，但通过对浓缩种子股票进行连续稀释，可以控制滴中生长的晶体数量。

使用 Seed Bead 套件，可以为后续的播种实验创建晶体种子储备。将晶体与种子珠一起放入微量离心管中，然后涡旋或超声以产生种子储备。然后通过进行连续稀释，可以控制实验中晶体的数量和大小。

Seed Bead 试剂盒可用于制备用于自动和半自动微播种的种子储备 (D’Arcy 2007, Harlos 2008)。

每个 Seed Bead Kit (HR2-320) 包含 24 个特殊的微量离心管，每个管包含一个 3mm PTFE Seed Bead。

每个 Seed Bead Steel Kit (HR4-780) 包含 24 个特殊的微量离心管，每个离心管包含 6 个 1.6 mm 不锈钢 Seed Beads。

每个 Seed Bead Ceramic Kit (HR4-781) 包含 24 个特殊的微量离心管，每个离心管包含 10 个 1.0 mm 硅酸锆陶瓷种子珠。

每个 Seed Bead Glass Kit (HR4-782) 包含 24 个特殊的微量离心管，每个管包含 10 个 1.0 mm 玻璃种子珠。

PTFE – Steel – Ceramic – Glass

 

Seed Bead Kit

Seed Bead Steel Kit

Seed Bead Ceramic Kit

Seed Bead Glass Kit

 HR2-320 Seed Bead User Guide HR4-780 Seed Bead Steel User Guide HR4-781 Seed Bead Ceramic User Guide HR4-782 Seed Bead Glass User Guide

1. Stura, E.A., Wilson, I.A., Methods: A Companion to Methods in Enzymology (1990) 1, 38-49.

2. Stura, E.A., Wilson, I.A., “Seeding Techniques” in Crystallization of Nucleic Acids and Proteins: A Practical Approach. Oxford University Press (1992) 99-126.

3. A method to produce microseed stock for use in the crystallization of biological macromolecules Luft, J.R., DeTitta, G.T. Acta Cryst. (1999). D55, 988-993.

4. J.R. Luft and G.T. DeTitta, Methods in Enzymology (1997) 276, 110-131.

5. Structure of an orthorhombic form of xylanase II from Trichoderma reesei and analysis of thermal displacement. Watanabe et al. Acta Cryst. (2006). D62, 784-792.

6. Crystallization and preliminary crystallographic analysis of p40phox, a regulatory subunit of NADPH oxidase. K. Honbou, S. Yuzawa, N. N. Suzuki, Y. Fujioka, H. Sumimoto and F. Inagaki. Acta Cryst. (2006). F62, 1021-1023 (Used seed bead to optimize)

7. Purification, crystallization and preliminary X-ray diffraction study of human ribosomal protein L10 core domain. Yuji Kobayashi et al. Acta Cryst. (2007). F63, 950-952

8. An automated microseed matrix-screening method. Allan D′Arcy, Frederic Villard and May Marsh. Acta Cryst. (2007). D63, 550-554

9. The role of bias in crystallization conditions in automated microseeding. Edwin Pozharski et al. Acta Cryst. (2008). D64, 1222-1227.

10. Transmission electron microscopy for the evaluation and optimization of crystal growth. Hilary P. Stevenson & Guillermo Calero et al. Acta Cryst. (2016) D72, 603-615.

11. Microseed matrix screening for optimization in protein crystallization: what have we learned? D’Arcy A, Bergfors T, Cowan-Jacob SW, Marsh M. Acta Crystallogr F Struct Biol Commun. 2014 Sep;70 (Pt 9):1117-26. doi: 10.1107/S2053230X14015507. Epub 2014 Aug 29.