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.<
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. RNeasy Protect Mini, Midi, and Maxi Kits2
. miRNeasy Mini Kit and miRNeasy 96 Kit