Easy Extraction of High Purity RNA from Cell, Tissue,and Yeast Samples Using Magnetic Silica Beads - No Phenol Extraction

MagExtractor® -RNA-


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This kit is for the extraction of high purity total RNA from biological samples such as cultured cells and tissues.
The recovered total RNA can be used mainly in enzymatic reactions such as RT-PCR.
As this kit employs the principle that magnetic silica beads bind total RNA present in a lysate solution, it is not necessary to perform deproteinization using harmful reagents such as phenol, ethanol precipitation, or high-speed centrifugation, which are necessary using the conventional AGPC method, thus simplifying the extraction process.
This kit is suitable for the MFX Series automatic nucleic acids extraction system and can also be used as a manual kit for B/F (solid-liquid) separation using a magnetic beads separation stand.

 

Application

 

  • Extraction of Total RNA from Cultured Cells, Tissue, and Yeast, etc.
    The recovered total RNA can be used in enzymatic reactions such as RT-PCR.

1. Examination of Yield and Purity
Total RNA was extracted from 2 x 106 HeLa cells using this kit and the widely
used AGPC RNA extraction method.
The yield obtained with this kit was approximately equivalent to that obtained
by AGPC method.
With regard to purity, the electrophoretic patterns and OD ratios indicate that
this kit extracts high purity RNA with almost no contamination by impurities
such as proteins and DNA.

2. Example PCR with Extracted RNA
1 µg of the total RNA extracted from 2 x 106 HeLa cells was used to perform RT-PCR with
transferrin receptor mRNA as a target (2115 bp).
As a result, amplification of the target was detected only in the reverse transcription reactions,
and expression of the normally low-volume expression transferrin receptor gene was confirmed.
The result showed that using RNA obtained with this kit enables RT-PCR with various targets.

3. Extraction from Rat Tissue
Total RNA was extracted from rat tissue using this kit.
50-100 mg of rat tissue was frozen in liquid nitrogen and crushed with a hammer. 750-900 µl of ice-cold Lysis & Binding Solution (2-ME content) was added, and the mixture homogenized completely with a pestle. The supernatant was removed and left at room temperature for 10 minutes to extract the total RNA. The results showed that by performing homogenization, using this kit enabled the extraction of high quality RNA, even from tissue.

 

Feature and Advantages

 

  • Suitable for Various Samples
    Allows extraction of total RNA from cultured cells, tissues, and yeast, etc.
    Total RNA mainly contains rRNA and mRNA.
  • Quick, Simple Extraction
    MagExtractor® -RNA- is based on the principle that magnetic silica beads bind total RNA, enabling quick simple extraction.
  • No Phenol or Chloroform Extraction
    This kit does not require the use of harmful phenol or chloroform for deproteinization so there is no hazardous waste problem.
  • Produces High Purity Total RNA
    The total RNA extracted with this kit rarely contains impurities such as DNA or proteins, allowing for use in enzymatic reactions such as RT-PCR.

 

Procedure

 

This kit extracts high purity total RNA according to the following steps.

  1. Add Lysis & Binding Solution containing chaotropic reagents to the sample to dissolve the cells.
  2. Add Magnetic Silica Beads and mix for the RNA in the sample to adhere to the silica's surface.
  3. Wash the beads with the two kinds of Washing Solution and remove unwanted proteins, DNA, salts, etc.
  4. Elute and recover RNA from the beads with sterile water or low salt buffer, etc.

 

One-Point Advice

 

  • B/F Separation
    Manual use of this kit is simplified with a magnetic beads separation stand.
    B/F separation may be performed using a compact desktop centrifuge instead of a magnetic beads separation stand.
  • Extraction of RNA from Blood
    The extraction of RNA from whole blood requires procedures such as Ficoll centrifugal separation of white blood cells, etc.
  • Sample Amount for Extraction
    Excessive sample amounts may result in reduced total RNA yields.
    We recommend the proper amount of sample be used.
  • Extraction of RNA from Tissue
    The extraction yield of RNA from tissue is different depending on the tissue used.
    Yield may be reduced when extracting from heart and muscle.
  • DNase I Treatment
    DNase I treatment prior to RT-PCR provides more stable results.

 

Product List

Product Name Cat# Quantity Price

MagExtractor -RNA- DataSheet ,

TYB-NPK-201F

100 RXN ¥36,100
$482

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To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.


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