Genomic DNA contamination is a major challenge in the RNA purification process.The most common method to remove genomic DNA contaminates is to use DNase digestion. The EZgene™ 96 HP RNA Isolation Kits are designed for fast isolation of total cellular RNA in high- throughput format without DNase I digestion. By using a special filter plate, genomic DNA can be effectively removed. The whole protocol can be completed in less than 40 minutes RNA purified using the EZgene™ DNA/RNA method is ready for applications such as RT-PCR, qPCR, differential display, microarrays, etc.
Principle
The EZgene™ RNA Isolation kit combines the reversible binding properties of RNA technology with a specially designed buffer system which can effectively remove DNA before RNA isolation. Samples are first lysed and homogenized in a specially designed denaturing buffer (LCT), which immediately inhibits the activity of RNase and DNase. The lysate is then passed through a gDNA Clearance Plate which traps the genomic DNA. After adjusting the binding condition, the flow-through lysate that contains RNA are bound to the RNA plate. After three wash steps, purified RNA is eluted with RNase-free water.
Storage and Stability
All components of the 96 Total RNA Kit plus should be stored at 22°C-25°C. Under these conditions, RNA has successfully been purified and used for RT-PCR after 12 months of storage. Under cool ambient conditions, precipitation may form in Buffer LY and RB Wash Buffer. The crystals may be dissolved by heating the buffer at 37°C.
Kit Contents
Catalog#
BMG-R6811-00
BMG-R6811-01
BMG-R6811-02
96 RNA Plate
1
4
12
96 DNA Clearance Plate
1
4
12
2 mL Deep Well Plate
2
4
24
500 µL 96 Well Plate
1
4
12
Buffer LY
20 mL
125 mL
400 mL
RB Wash Buffer
70 mL
280 mL
850 mL
RNA Wash Buffer
40 mL
160 mL
2 x 200 mL
DEPC Water
10 mL
40 mL
120 mL
User Manual
1
1
1
Preparing Reagents
Dilute RNA Wash Buffer with absolute ethanol (96-100%) as follows
Kit
Ethanol to be added
BMG-R6811-00
Add 160 mL absolute ethanol
BMG-R6811-01
Add 800 mL absolute ethanol
BMG-R6811-02
Add 800 mL absolute ethanol per bottle
Materials to be provided by user
96%-100% ethanol
70% ethanol • Multichannel pipette
RNase-free filter pipette tips
Reagent reservoirs for multichannel pipettes
Centrifuge with rotor for 96-well plates
Disposable latex gloves
2ml 96-well deep well plate
Sealing film
Swing Bucket Centrifuge capable of 5,000 x g and Adaptor for 96 Deep Well plates
