miRNA Isolation Sample Kit – 4 Preps
- Sample Size: 100mg tissue/1×106 cultured cells
- Operation Time: 30min or less
- Elution Volume: 50µl
- Free sample available upon request
The miRNA Isolation Kits are designed for the purification of microRNA (miRNA) and other small cellular RNAs from tissue samples and cultured cells. Purification of miRNA allows research into significant biological pathways for gene regulation. The standard protocol for isolating total RNA and miRNA are not optimized for small RNA molecules, and therefore result in the loss of substantial amounts of miRNA and other small RNA. In addition, the removal of the predominantly larger RNAs is required for accurate analysis of miRNA expression by qPCR or microarray analysis. These kits are specifically designed for the purification of small RNA with minimal contamination from large RNA molecules or genomic DNA. The method employs a spin column with a silica-based filter matrix that binds RNA in the presence of a chaotropic salt. This method is based on the selective binding of RNA molecules of various sizes to the silica-based fiber matrix when different ethanol concentrations are present in the solvent.
|Specifications||miRNA Isolation Kit|
|Binding Capacity:||50 µg|
|Sample Concentration:||100 mg of Tissue, up to 1 x 106 Cultured cells|
|Sample Type:||Animal tissue, cells|
|Typical Yield:||5-30 µg|
|Elution Volume:||50 µl|
|Operation Time:||<30 mins.|
What is the binding capacity of the miRNA Isolation column? The binding capacity is approximately 30 µg.
What are the comparable competitor’s kits? QIAGEN miRNeasy Mini Kit
Can replacement columns used for Total RNA also be used for miRNA? No, there are subtle differences between the Total RNA columns and miRNA columns.
Can the miRNA kit be used to isolate miRNA from urine? Yes, these initial steps must be followed. 1. Transfer 10 ml of urine into a 15 ml centrifuge tube (RNAse free) and centrifuge at 6,000 x g for 2 minutes. 2. Discard the supernatant; add 500 µl of RNAse Free Water to the pellet and vortex for 5 seconds. 3. Transfer the sample into a 1.5 ml micro-centrifuge tube (RNAse free). 4. Centrifuge at 6,000 x g for 2 minutes, and then discard the supernatant. Follow the miRNA Isolation kit protocol step 1 Lysis. Add 200 µl of Lysis Buffer into a 1.5 ml micro-centrifuge tube containing the cell pellet.
What is the make up of the Lysis buffer? The Lysis buffer for the miRNA isolation kit consists of Guanidinium Isothyocyanate and a detergent – sodium lauroyl sarcosinate.
Does the Lysis buffer merely breakdown the cell membrane to access the miRNA or does it lyse the entire cell nucleus? The Lysis buffer in the miRNA isolation kit will lyse the entire cell nucleus.
What is the size of the purified RNA’s from this miRNA kit? Most of the RNA purified using the miRNA kit are less than 100 nt. Slightly larger RNA’s can be also be purified by using a modified protocol.