For total RNA isolation from a wide variety of sources
RNeasy Protect Midi Kit
RNeasy Protect Maxi Kit
Mini columns with 1.5 ml and 2 ml collection
Midi columns with 15 ml collection tubes
Maxi columns with 50 ml collection tubes
150 mg 1 g
50 ml (50-prep
250 ml (250-prep kit)
20 ml (10-prep
100 ml (50-prep kit)
100 ml (12-prep
Up to 100 g
Up to 1 mg
Up to 6 mg
on sample source. See table
RNeasy Mini, Midi, and Maxi Kits allow efficient purification
of total RNA from very small (see figure "RT-PCR of
RNA from =100 Cells
") to very large amounts of starting material.
Total RNA is easily purified from animal cells or tissues, Gram-positive
or Gram-negative bacteria,* or yeast (See table "Total
RNA Yields Obtained with RNeasy Mini, Midi, and Maxi Kits
figure "High-Quality RNA from a Variety of Samples
Protocols are also included for isolation of cytoplasmic RNA from
eukaryotic cells as well as for cleanup of partially purified RNA, in vitro
transcripts, and RNA from enzymatic reactions. A variety of special application
protocols, including a protocol for vacuum processing of RNeasy mini columns,
are also available. QIAshredder Homogenizers for convenient cell-lysate
homogenization are available as an accessory.
RNeasy Kits provide the highest-quality RNA with minimum copurification
of DNA. For certain RNA applications that are sensitive to very small
amounts of DNA, the residual amounts of DNA remaining can be removed using
the RNase-Free DNase Set for convenient on-column DNase treatment during
the RNeasy procedure.
* Lysozyme (required for bacterial samples) is not provided.
High-Quality RNA from a Variety of Samples
Lyticase, zymolase, or glass beads (required for yeast samples)
are not provided.
Additional buffer required. The buffer composition is provided
in the protocols.
Formaldehyde agarose ge
l and northern blot
of total RNA purified with the RNeasy Maxi Kit. 10 g of total RNA
isolated from each source was loaded per lane. All tissues were from mouse.
Yeast: Saccharomyces cerevisiae; E. coli: HB101. 32P-labeled probes recognized
(G) GAPDH; (E) translation elongation factor EF-1a ; and (O) outer membrane
protein A sequences. (E and O were kindly provided by P. Philippsen, University
of Basel, Switzerland and U. Henning, Max Planck Institute of Biology, Tbingen,
Germany, respectively.) B. subtilis was not probed. M: 0.249.5 kb
RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes
is a nuclear precursor RNA.
RT-PCR of RNA from >100 Cells
RT-PCR of total RNA isolated with the RNeasy Mini Kit
from the indicated numbers of HeLa cells. 10 l (1/5) of eluate was
digested with RNase-free DNase and reverse transcribed with oligo-dT primer.
2.5 l (1/20) of the cDNA mix was used in 50 l PCR. A 452 bp
fragment of GAPDH was amplified. C: negative control; C+: positive
control; M: 100 bp ladder.
Total RNA Yields Obtained with RNeasy Mini, Midi, and Maxi
Amounts can vary due to developmental stage, species, growth
conditions used, etc. Since the RNeasy procedure enriches for RNA species
>200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight
* Using the specialized protocol for heart, muscle, and skin tissue, included
in handbook. Proteinase K (required for this protocol) is not provided.
Higher yields can be obtained from stabilized bacteria using RNeasy
Protect Bacteria Kits.<
* Requires use of a centrifuge capable of attaining 30005000 x g equipped
with a swing-out rotor for 15 ml (Midi) or 50 ml (Maxi) centrifuge tubes.
Source:Page: All 1 2 3 4 Related biology technology :1
. RNeasy Protect Mini, Midi, and Maxi Kits2
. miRNeasy Mini Kit and miRNeasy 96 Kit