MagExtractor®-RNA-
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.
- Add Lysis & Binding Solution containing chaotropic reagents to the sample to dissolve the cells.
- Add Magnetic Silica Beads and mix for the RNA in the sample to adhere to the silica's surface.
- Wash the beads with the two kinds of Washing Solution and remove unwanted proteins, DNA, salts, etc.
- 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.
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MagExtractor -RNA- | TYB-NPK-201 | 100 RXN | €320 |