Purification of total RNA using the Marligen Total RNA Rapid Purification System
Purification of total RNA using the Marligen Total RNA Rapid Purification System
The Rapid Total RNA System by Marligen Biosciences extracts RNA from a variety of samples with very consistent yield and quality. This experiment was derived from PCR Laboratory Guide (Second Edition) by Kang Seed and Lijia Qu.
Operation method
Purification of total RNA using the Marligen Total RNA Rapid Purification System
Materials and Instruments
Ethanol β-mercaptoethanol Move I. Materials For more product details, please visit Aladdin Scientific website.
Rotor-stator homogenizer or similar device Marligen Rapid Total RNA System
1. buffers, solutions and reagents
Ethanol, 70% and 100%.
β-mercaptoethanol
2.Special equipment
Rotor-stator homogenizer or similar equipment
3.Other
Marligen Rapid Total RNA System (Marligen Biosciences)
II. Methods
(1) Prepare a lysis solution by adding β-mercaptoethanol.
(2) Add lysis solution to cell and tissue samples according to the manufacturer's protocol (volume of lysate depends on the size of the sample being processed). Homogenize with a rotor-stator homogenizer or similar device.
(3) Add an equal volume of 70% ethanol to the lysis product. Homogenize the lysis product to disperse the precipitate.
(4) Place the lysis product in the centrifugal sleeve column provided with the kit and centrifuge at 12,000 to 16,000 g in a microcentrifuge.
(5) Wash once with 700ul of wash buffer 1 and centrifuge at 12000~16000g for 1min.
(6) Wash twice with washing buffer 2, 500ul each time, and centrifuge at 12000~16000g for 1min.
(7) Centrifuge at 12000~16000g for 2min to remove the residual wash buffer.
(8) Elute RNA from the column with appropriate water according to the instructions of the kit and store at -70°C. For small samples, elute with 45ul of water; for large samples, elute with 20ul of water. 