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).
引用および参考文献 (6)
引用および参考文献
Abstract
[Differential expression profile of microRNA between hyperplastic scar and normal skin].
Authors:Ning P, Liu DW, Mao YG, Peng Y, Lin ZW, Liu DM,
Journal:Zhonghua Yi Xue Za Zhi
PubMed ID:22781298
To explore the miRNA differential expression profiles of hyperplastic scar and normal skin so as to further elucidate the pathogenesis of hyperplastic scar and search for new therapeutic targets. The total RNA was extracted from 5 human hyperplastic scar and normal skin tissues by Trizol. The specimens were collected from ... More
Circulating microRNAs in serum: novel biomarkers for patients with bladder cancer?
Authors:Scheffer AR, Holdenrieder S, Kristiansen G, von Ruecker A, Müller SC, Ellinger J,
Journal:World J Urol
PubMed ID:23266581
'PURPOSE: Recent studies indicate that circulating microRNAs in serum/plasma are a novel class of non-invasive biomarkers with diagnostic and prognostic information. So far, circulating microRNAs have not been analyzed in patients with bladder cancer. METHODS: We collected serum from patients with non-muscle invasive bladder cancer (NMIBC), muscle invasive bladder cancer ... More
[Isolation, identification and analysis of the expression profile of miRNAs in Aedes albopictus].
Authors:Zheng PM, Wu JY, Gu JB, Tu ZJ, Chen XG,
Journal:Nan Fang Yi Ke Da Xue Xue Bao
PubMed ID:20423824
'To verify the miRNA in Aedes albopictus and characterize the expression profile of several miRNAs across all the life stages of Aedes albopictus. Based on the published miRNA sequences of Anopheles gambiae and Aedes aegypti, 6 DIG-labeled antisense probes were synthesized. The total RNAs from Aedes albopictus in 6 developmental ... More
MicroRNA expression profiles in the progression of prostate cancer-from high-grade prostate intraepithelial neoplasia to metastasis.
Authors:Leite KR, Tomiyama A, Reis ST, Sousa-Canavez JM, Sañudo A, Camara-Lopes LH, Srougi M,
Journal:Urol Oncol
PubMed ID:21880514
'Models of the multistep process related to cancer progression have been designed for many cancers including prostate. The aim of this study is to propose a new model including a possible role for recently described micro RNAs in prostate cancer (CaP) progression. Sixty-three patients underwent radical prostatectomy to treat localized ... More
A model for data analysis of microRNA expression in forensic body fluid identification.
Authors:Wang Z, Luo H, Pan X, Liao M, Hou Y,
Journal:Forensic Sci Int Genet
PubMed ID:21903498
MicroRNAs (miRNAs, 18-25 bases in length) are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. MiRNA expression patterns, including presence and relative abundance of particular miRNA species, provide cell- and tissue-specific information that can be used for body fluid identification. Recently, two published studies reported that a ... More