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mirVana™ miRNA Isolation Kit, with phenol - FAQs

查看更多产品信息 mirVana™ miRNA Isolation Kit, with phenol - FAQs (AM1560)

7 个常见问题解答

对于从组织、细胞或液体样本中分离miRNA，您建议使用什么试剂盒?

对于组织或细胞，我们推荐mirVana miRNA分离试剂盒(货号AM1560)。对于液体样品，我们推荐mirVana Paris RNA和天然蛋白纯化试剂盒(货号AM1556)。对于高通量miRNA分离，我们建议使用MagMAX mirVana总RNA分离试剂盒(货号A27828)< / p >

纯化miRNA样本的不同方法有哪些？

沉淀miRNA：根据mirVANA miRNA Isolation试剂盒或mirVANA PARIS试剂盒的说明进行操作。
两次酚/胍(离液试剂)提取：使用酚/胍盐裂解组织，然后用酚提取，离心，取上清，并沉淀RNA。
玻璃纤维过滤：使用离液剂裂解组织，将RNA结合在玻璃纤维膜上，然后洗涤除去未结合的DNA/蛋白/盐/核苷酸，然后洗脱RNA。

What kit would you recommend for isolation of miRNA from tissue, cells, or liquid samples?

For tissue or cells, we recommend the mirVana miRNA Isolation Kit (Cat. No. AM1560), while for liquid samples, we recommend the mirVana Paris RNA and Native Protein Purification Kit (Cat. No. AM1556). For high-throughput miRNA isolation we recommend using the MagMAX mirVana Total RNA Isolation kit (Cat. No. A27828)

What are different methods for cleaning up my miRNA sample?

- Precipitation of miRNA: Follow the instructions for the mirVANA miRNA Isolation Kit or the mirVANA PARIS RNA and Native Protein Purification Kit.
- Double phenol/guanidium (chaotropic reagent) extraction: Disrupt tissue with phenol/guanidine, then extract with phenol, centrifuge, remove the supernatant, and precipitate the RNA.
- Glass-fiber filter: Disrupt the tissue in a chaotropic agent, bind the RNA to a glass-fiber filter, then wash off unbound DNA/protein/salt/nucleotides and elute the RNA.

What type of sample can be used with TaqMan MicroRNA Assays?

One to ten nanograms of total RNA per 15 µL RT reaction is recommended for use with the TaqMan MicroRNA Assay. We have successfully detected miRNAs in as little as 25 pg of total RNA (~100 cells) in samples where the miRNA was highly abundant. The total RNA isolation procedure used must preserve small RNA species. We recommend the Invitrogen mirVana miRNA Isolation Kit (Cat. No. AM1560) or TRI reagent (Cat. No. AM9738).

When using the mirVana miRNA Isolation Kit, with phenol (Cat. No. AM1560), can I store homogenized tissue RNA lysate in the lysis buffer at -80 degrees C?

Yes, the RNA lysates can be stored at -80 degrees C for up to 2 months.

What kit do you recommend to capture small RNA that is >17 nt in size?

We recommend using the mirVana miRNA Isolation Kit (Cat. No. AM1560, AM1561), both of which are designed to capture small RNAs, such as miRNAs, tRNAs, etc. Purification of RNAs as small as 20 nt has been obtained in-house using these kits. Note: Cat. No. AM1561 does not contain phenol, but is required for the procedure. Phenol can be purchased separately (Cat. No. AM9720).