Bangs Laboratories 提供多种用于微粒试剂开发的附件产品。该系列包括交联剂和表面活性剂，以及用于微球涂层和储存的溶液和缓冲液。这些产品补充了我们广泛的聚合物、二氧化硅和磁性微粒目录，还提供了一种方便的方法来补充偶联试剂盒的各个组件。
Product Data Sheet 530 BioMag® Streptavidin,Nuclease-free
Description
BioMag® Streptavidin is a nuclease-free suspension of BioMag® particles
approximately 1.5µm in size, which are covalently coated with streptavidin.
The suspension is supplied in a phosphate buffered saline (pH 7.4)
containing 0.1% BSA. Sodium azide has been added as an anti-microbial.
Shake vigorously or vortex before use. Magnetically separate the BioMag®
particles, aspirate the supernatant, and resuspend in an appropriate buffer.
BioMag® Streptavidin 是 BioMag® 颗粒的无核酸酶悬浮液大小约为 1.5 µm，共价包被链霉亲和素。
悬浮液以磷酸盐缓冲盐水 (pH 7.4) 形式提供
含 0.1% BSA。 已添加叠氮化钠作为抗微生物剂。
使用前用力摇晃或涡旋。 磁性分离 BioMag®颗粒，吸出上清液，并重悬于适当的缓冲液中。
Characteristics
Mean Diameter: ~1.5µm
Particle Concentration: 1mg/mL
Binding Capacity: 1mg of BioMag® Streptavidin will bind:
>1500 pmoles of free biotin
>1000 pmoles of a 20-mer biotinylated
oligonucleotide
>200 pmoles of a 100-mer biotinylated
oligonucleotide
>70 pmoles of a 300 bp 5-biotinylated double
stranded DNA
>25 pmoles of a 1Kbp 5-biotinylated double-s
tranded DNA
Material
Material Supplied
• BioMag® Streptavidin: 10mL or 25mL
Material Required
• Binding Buffer: 20mM Tris and 0.5M NaCl at pH 8.0
• Wash Buffer: 7mM Tris and 0.17M NaCl at pH 8.0
• DEPC-treated water
• Nuclease-free microcentrifuge tubes
• Magnetic separator
Procedure
Researchers are advised to optimize the use of BioMag® in any application
as procedures designed by other manufacturers may not be ideal.
The following procedure is for the isolation of 1-2µg of mRNA from
approximately 75-100µg of total RNA. The total isolation time is less than 30
minutes.
1. Dispense 200µL of BioMag® Streptavidin into a nuclease-free
microcentrifuge tube. Using a magnetic separation unit, pull the
magnetic particles to the side of the microcentrifuge tube for 30
seconds. Remove and discard the supernatant. Resuspend the
BioMag® Streptavidin in 100µL of Binding Buffer.
2. Incubate 2.5µL (2.5µg) of 5-Biotinylated Oligo (dT) (or an appropriate
amount of biotinylated molecule) with the 100µL of BioMag®
Streptavidin from Step 1 for 15 minutes at room temperature.
3. Magnetically separate for 30 seconds and discard the supernatant.
Wash the Oligo (dT) bound particles from Step 2 with 100µL of Binding
Buffer 2 times, leaving the magnetic particles as a wet cake.
4. Bring up the total RNA sample in DEPC-treated water to a total volume
of 90µL.
5. Incubate the RNA sample at 55˚C for 5 minutes to disrupt secondary
structures.
6. Add 10µL of 5M NaCl to achieve a final concentration of 0.5M NaCl.
7. Add the total RNA to the washed magnetic particles from Step 3. Mix
gently and hybridize at room temperature for 3 minutes.
8. Magnetically separate and wash the particles with 100µL of Wash
Buffer.
9. Elute the bound mRNA with 25-50µL of DEPC-treated water at 55˚C for
2 minutes.
10. Magnetically separate and transfer the supernatant to a nuclease-free
microcentrifuge tube.
11. Repeat elution of mRNA with 25-50µL of DEPC-treated water at 55˚C
for another 2 minutes in order to completely elute the bound mRNA
from the particles. Magnetically separate and transfer the supernatant
to the tube containing the first elution of mRNA from Step 10.
建议研究人员在任何应用中优化 BioMag® 的使用因为其他制造商设计的程序可能并不理想。
以下程序用于从大约 75-100 µg 的总 RNA。总隔离时间小于30分钟。
1. 将 200µL BioMag® Streptavidin 分装到无核酸酶中微量离心管。使用磁性分离装置，拉动将磁性颗粒移至微量离心管侧面 30秒。取出并丢弃上清液。重新挂起
BioMag® Streptavidin 在 100µL 结合缓冲液中。
2. 孵育 2.5µL (2.5µg) 5-生物素化寡核苷酸 (dT)（或适当的生物素化分子的量）与 100µL BioMag®步骤 1 中的链霉亲和素在室温下放置 15 分钟。
3. 磁力分离 30 秒，弃去上清液。用 100µL Binding 清洗步骤 2 中的 Oligo (dT) 结合颗粒缓冲 2 次，留下磁性颗粒作为湿饼。
4. 将 DEPC 处理水中的总 RNA 样品调至总体积90µL。
5. 将 RNA 样品在 55˚C 下孵育 5 分钟以破坏二代结构。
6. 加入 10µL 5M NaCl 以达到 0.5M NaCl 的最终浓度。
7. 将总 RNA 添加到步骤 3 中清洗过的磁性颗粒中。混合
轻轻地在室温下杂交 3 分钟。
8. 用 100µL Wash 磁力分离和清洗颗粒缓冲。
9. 用 25-50µL DEPC 处理的水在 55˚C 洗脱结合的 mRNA2分钟。
10. 磁性分离上清液并将其转移至无核酸酶微量离心管。
11. 用 25-50µL DEPC 处理水在 55˚C 重复洗脱 mRNA再过 2 分钟以完全洗脱结合的 mRNA从粒子。磁性分离和转移上清液到第 10 步中第一次洗脱 mRNA 的试管。
References
1. Hornes, E., L. Korsnes. 1990. Magnetic DNA hybridization properties
of oligonucleotide probes attached to superparamagnetic beads and
their use in the isolation of poly(A) mRNA from eukaryotic cells. Genet
Anal Tech Appl; 7(6):145-150.
2. Morrissey, D.V., M. Lombardo, J.K. Eldredge, K.R. Kearney,
E.P. Groody, M.L. Collins. 1989. Nucleic acid hybridization assays
employing dA-tailed capture probes. Multiple capture methods. Anal
Biochem, 181(2):345-359.
Storage and Stability
Store at 2-8˚C. Freezing, drying, or centrifuging BioMag® may result in
irreversible aggregation and loss of binding activity.
储存于 2-8°C。 冷冻、干燥或离心 BioMag® 可能会导致不可逆的聚集和结合活性的丧失。
Safety
This particle suspension contains sodium azide. Sodium azide may react with
lead and copper plumbing to form explosive metal azides. Upon disposal of
material, flush with a large volume of water to prevent azide accumulation.
Please consult the Material Safety Data Sheet for more information.
These products are for research use only and are not intended for
use in humans or for in vitro diagnostic use.
这种颗粒悬浮液含有叠氮化钠。 叠氮化钠可能与铅和铜管道形成爆炸性金属叠氮化物。 处置后材料，用大量水冲洗以防止叠氮化物堆积。
请查阅材料安全数据表了解更多信息。这些产品仅供研究使用，不适用于用于人类或体外诊断用途。
Ordering Information
Cat. Code Description Size
BM568 BioMag® Streptavidin, Nuclease-free 10mL or 25mL