digest/homogenize with Trizol. Use the miRneasy Kit from Qiagen

You can directly go for usual TriZol method.

For breast cancer cells miRNA isolation, we isolate total RNA, then convert all present miRNA to cDNA for gene expression. Total RNA method works wonderfully, since we tried with Thermo kit previously but Yield issue was coming.

Use miRVana - miRNA isolation kit from Thermo Fisher Scientific

Thanks all for the help!

Hi Selina,

Since your target is to isolate total RNA including small RNA, you should be using the Animal Tissue RNA Purification Kit (Cat. 25700) from Norgen Biotek. Unlike Qiagen’s or miRVana, Norgen’s kit doesn’t use phenol/chloroform for the tissue RNA extraction and therefore the recovered RNA is inhibitor-free without any possible carry-over contaminants especially for your RNAseq which is a very sensitive downstream application. Additionally, Norgen’s uses its proprietary Silicon Carbide spin column which differs from the Silica spin columns used in other kits (miRneasy and miRVana). Unlike silica-based RNA isolation, Norgen’s Silicon Carbide binds all RNA sizes from large mRNA and ribosomal RNA down to microRNA (miRNA). Also silica based columns tends to have a bias towards binding small RNA sequences with high GC content whereas Norgen’s Silicon Carbide matrix binds all small RNA irrespective of GC content. Please check the following links to see some of the data showing the differences between Silica and Norgen’s silicon Carbide.

https://norgenbiotek.com/sites/default/files/resources/microRNA-2009-Poster.pdf

https://norgenbiotek.com/sites/default/files/resources/total_rna_purification_kit_variation_between_the_purification_efficiency_of_microrna_with_different_gc_content_in_biological_and_clinical_specimens_posters_68.pdf

Thanks very much, that's very useful information!