流式细胞仪-Flow Cytometry

你好Flow-ers、Flowbos、Cytometrians，或者只是流式细胞仪爱好者。欢迎来到我们的流式细胞仪部门！我们很长一段时间都在研究珠子，并且很自豪能够延续我们的前身流式细胞术标准公司开始的流式细胞术标准化传统。我们提供范围广泛的产品，支持仪器验证和 QC，以及分析和多站点标准化。

Product Data Sheet 529 BioMag® Oligo dT(20),Nuclease-free

Description

BioMag® Oligo dT(20) is a nuclease-free suspension of BioMag® particles
approximately 1.5µm in size, which are covalently bound with Oligo dT(20).
The suspension is supplied in a buffer (pH 8.0) containing 20 mM Tris,
0.5 M sodium chloride, and 0.1% sodium azide. Shake vigorously before
use. Magnetically separate the BioMag® particles, aspirate the supernatant,
and resuspend in an appropriate buffer for your application.

BioMag® Oligo dT(20) 是 BioMag® 颗粒的无核酸酶悬浮液大小约为 1.5µm，与 Oligo dT(20) 共价结合。悬浮液在含有 20 mM Tris 的缓冲液 (pH 8.0) 中提供，
0.5 M 氯化钠和 0.1% 叠氮化钠。之前用力摇晃采用。磁性分离 BioMag® 颗粒，吸出上清液，并重新悬浮在适合您的应用程序的缓冲区中。

Characteristics

Mean Diameter: ~1.5µm
Particle Concentration: 5mg/mL
Binding Capacity: 100µL of BioMag® Oligo dT(20) is sufficient to
isolate ~1-2µg of polyadenylated RNA from
~100µg of total RNA

平均直径：~1.5µm
颗粒浓度：5mg/mL
结合能力：100µL BioMag® Oligo dT(20) 足以
从
~100µg 总 RNA

Material

Material Supplied
• BioMag® Oligo dT(20): 2mL
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 is a procedure for the isolation of 1-2µg of polyadenylated RNA
from approximately 100µg of total RNA. The 2mL of BioMag® Oligo dT(20)
supplied is sufficient for 20 isolations of 1-2µg of mRNA. (More or less
mRNA can be isolated by modifying the procedure.) The total isolation time is
approximately 15 minutes.
1. Dispense 100µL of BioMag® Oligo dT(20) 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. Wash the BioMag®
Oligo dT(20) once with 200µL of the Binding Buffer. Magnetically
separate, discard the supernatant, and resuspend in 100µL of the
Binding Buffer.
2. Bring up the total RNA sample in DEPC-treated water to a total volume
of 90µL.
3. Incubate the RNA sample at 55˚C for 5 minutes to disrupt secondary
structures.
4. Add 10µL of 5M NaCl to achieve a final concentration of 0.5M NaCl.
5. Add the total RNA to the BioMag® Oligo dT(20) from Step 1. Mix gently
and hybridize at room temperature for 3 minutes.
6. Magnetically separate and wash the particles 2 times with 100µL of
the Wash Buffer.
7. Elute the bound polyadenylated RNA with 25-50µL of DEPC-treated
water at 55˚C for 2 minutes. Greater than 90% of polyadenylated RNA
is eluted in this step.
8. Magnetically separate and transfer the supernatant to a nuclease-free
microcentrifuge tube.
9. Repeat elution of polyadenylated RNA 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 7.

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。
以下是分离 1-2µg 多聚腺苷酸化 RNA 的步骤
从大约 100 µg 的总 RNA 中提取。 2mL BioMag® Oligo dT(20)
提供的 1-2 µg mRNA 足以分离 20 次。（或多或少可以通过修改程序来分离 mRNA。）总分离时间为大约 15 分钟。
1. 将 100µL BioMag® Oligo dT(20) 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。清洗 BioMag®
Oligo dT(20) 与 200 µL 结合缓冲液一起使用。磁性的分离，弃去上清液，重悬于 100 µL绑定缓冲区。
2. 将 DEPC 处理水中的总 RNA 样品调至总体积90µL。
3. 将 RNA 样品在 55˚C 下孵育 5 分钟以破坏二代结构。
4. 加入 10µL 5M NaCl 以达到 0.5M NaCl 的最终浓度。
5. 将步骤 1 中的总 RNA 添加到 BioMag® Oligo dT(20) 中。轻轻混合并在室温下杂交 3 分钟。
6. 用 100µL 磁力分离和洗涤颗粒 2 次洗涤缓冲液。
7. 用 25-50µL 经 DEPC 处理的多聚腺苷酸化 RNA 洗脱55℃的水2分钟。大于 90% 的多聚腺苷酸化 RNA在该步骤中被洗脱。
8. 磁性分离并将上清液转移至无核酸酶微量离心管。
9. 用 25-50µL DEPC 处理的多聚腺苷酸 RNA 重复洗脱55˚C 的水再 2 分钟，以完全洗脱从颗粒中结合 mRNA。磁性分离和转移上清液到含有第一次洗脱的 mRNA 的试管中步骤 7。

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. To ensure stability,
BioMag® Oligo dT(20) must be stored in the buffer in which it is supplied.

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。为确保稳定性，
BioMag® Oligo dT(20) 必须储存在提供它的缓冲液中。

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
BM529 BioMag® Oligo dT(20), Nuclease-free 2mL