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EXTRACTME RNA BACTERIA & YEAST KIT

EXTRACTME RNA BACTERIA & YEAST KIT
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The EXTRACTME RNA BACTERIA & YEAST kit is designed for the rapid and efficient purification of high quality total RNA from broth yeast or bacteria cultures as well as frozen cells. The isolation protocol and buffer formulations were optimized for high isolation efficiency and purity of RNA. The product is intended for research use only.
 
SAMPLE MATERIAL
- broth bacterial (up to 1x 109 cells) or yeast (up to 5x 107 cells) culture
- frozen cell pellet
 
YIELD
It depends on the sample material amount and type.
- up to 60 μg RNA from bacterial culture
- up to 30 μg RNA from yeast culture
 
BINDING CAPACITY
approx. 90 μg RNA
 
TIME REQUIRED
- 30 minutes for RNA isolation from bacteria
- 50 minutes for RNA isolation from yeast
 
RNA PURITY
A260/A280 ratio = 1.9 – 2.1
 
Principle
The EXTRACTME RNA BACTERIA & YEAST kit utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids at high concentration of chaotropic salts. In the first step of the isolation procedure the cell walls are disintegrated and the membranes and proteins are degraded. Any RNases are inactivated by guanidine thiocyanate and β-mercaptoethanol. The homogenate is separated from the undigested cell remains by centrifugation and on the Homogenizing Column. The RNA is bound to the Purification Column membrane by addition of ethanol. The three-step washing stage effectively removes impurities and enzyme inhibitors. The purified RNA is eluted using a low ionic strength buffer or RNase-free water (pH 7.0-9.0) and can be used directly in all downstream applications such as RT-PCR, Northern blotting, RT-qPCR and so forth.
 
Quality control
The quality of each production batch (LOT) of the EXTRACTME RNA BACTERIA & YEAST kit is tested using standard QC procedures. The purified RNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by reverse transcription and qPCR.
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