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Cat. No.: BMRE-50 (for 50T)

Cat. No.: BMRE-200 (for 200T)

Description

Traditional microRNA extraction kits are typically based on the principle of using guanidinium isothiocyanate/phenol/chloroform (TRIzol method) combined with high-concentration ethanol to adsorb microRNA fragments onto silica membranes. However, complex bone tissues are hard, have low bone cell density, and their peripheral matrix contains large amounts of mucoproteins (glycoproteins) that make RNA separation challenging. This makes it impossible to extract high-quality RNA/microRNA using the traditional TRIzol method. Consequently, most companies that rely on the TRIzol principle for their microRNA extraction kits, are at a loss when faced with bone tissue samples. Their product lines simply lack a dedicated bone tissue microRNA extraction kit. In desperation, researchers resort to traditional methods or TRIzol extraction, using isopropanol/ethanol precipitation to extract microRNA. While this might extract some microRNA, the precipitation method often results in significant losses or co-precipitation with impurities, compromising experimental results. When submitting to international journals, these results frequently face scrutiny, and some papers might even be retracted. This kit, built on our company's globally leading RNA/microRNA extraction technology, innovatively avoids the use of phenol and chloroform, making us the first in the world to address this issue. It allows for the extraction of microRNA from most bone tissue samples.

Our unique lysis solution swiftly disrupts cells and inactivates cellular RNAases. The plant RNA aid, PLANTaid, helps bind polysaccharides and polyphenols, which are then removed by centrifugation. Following this, the lysate is passed through a genomic DNA removal column. Genomic DNA is effectively cleared, while RNA (including microRNA) is selectively filtered. The filtered RNA is then conditioned with ethanol and, under high salt conditions, selectively adsorbs onto a silica membrane within the centrifuge column. Through a series of specialized washing steps followed by centrifugation, cellular metabolites and proteins are efficiently removed. Finally, using low-salt RNase-free H2O, the purified RNA (including microRNA) is eluted from the silica membrane.

Employing segmented extraction steps, it's also possible to separately purify the enriched microRNA fraction and total RNA (>200 nt), thus obtaining isolated and enriched microRNA and RNA.

Features

• Completely avoids the use of toxic reagents like phenol, chloroform, and does not require ethanol precipitation steps.
• Fast and simple, a single sample operation can generally be completed within 30 minutes.
• The unique RNA boosting agent effectively binds to proteoglycans, enhancing the purification process.
• Multiple column washes ensure high purity with a typical OD260/OD280 ratio of 1.9 to 2.0, suitable for RT-PCR, Northern-blot, and various other experiments.

Storage

This kit can be stored at room temperature for 6 months without affecting its performance.