Manual Part #740000-11 Revision #035004a
Copyright © 2005 by Stratagene
Universal Human Reference RNA
Catalog #740000
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Storage Store the Universal Human Reference RNA at –80°C. Store the RNase-free water at –20°C.
I
NTRODUCTION
Stratagene’s Universal Human Reference RNA is composed of total RNA from 10 human cell lines. The reference RNA is designed to
be used as a reference for microarray gene-profiling experiments. Since RNA species differ in abundance between cell lines, an ideal
reference sample should represent these different RNAs. Equal quantities of DNase-treated total RNA from each cell line were
pooled to make the Universal Human Reference RNA. This Universal Reference RNA is suitable for microarray experiments.
Stratagene also supplies a QPCR Human Reference Total RNA, suitable for QRT-PCR, which has undergone further DNase treatment.
M
ATERIALS
P
ROVIDED
Material Provided
Quantity
Reference RNA
2 tubes x 200 µg each
RNase-free water
1.5 ml
Cell Line Derivations
Adenocarcinoma, mammary gland
Melanoma
Hepatoblastoma, liver
Liposarcoma
Adenocarcinoma, cervix
Histiocytic lymphoma; macrophage; histocyte
Embryonal carcinoma, testis
Lymphoblastic leukemia, T lymphoblast
Glioblastoma, brain
Plasmacytoma; myeloma; B lymphocyte
A
DDITIONAL
M
ATERIALS
R
EQUIRED
RNase-free 70% Ethanol
P
ROTOCOL
Universal Human Reference RNA is provided in a solution of 70% ethanol and 0.1 M sodium acetate. Prepare the Reference RNA for
use as follows:
1. Centrifuge the tube at 12,000 × g for 15 minutes at 4°C.
2. Carefully remove the supernatant.
3. Wash the pellet in 70% ethanol.
4. Centrifuge the tube at 12,000 × g for 15 minutes at 4°C.
5. Carefully remove the supernatant and air-dry the pellet at room temperature for 30 minutes to remove retained ethanol.
6. Resuspend the pellet in RNase-free water to the desired concentration.
Proceed with the preparation of labeled cDNA and interrogate the arrays according to the manufacturer’s instructions.
Q
UALITY
C
ONTROL
T
ESTING
The quality of the Universal Human Reference RNA is assessed by observing distinct 28S and 18S ribosomal bands on a 1× MOPS
agarose gel under denaturing conditions. The purity of the RNA is assessed by spectrophotometry (A260/A280
≥1.8). The RNA is then
shown to be free of contaminating RNases by incubation in a suitable buffer at 37
°C followed by gel analysis against known
RNAse-free controls. The RNA is further tested functionally by synthesizing labeled cDNA, which is then hybridized to a microarray
to examine gene representation and coverage.
L
IMITED
P
RODUCT
W
ARRANTY
This warranty limits our liability to replacement of this product. No other warranties of any kind, express or implied, including without
limitation, implied warranties of merchantability or fitness for a particular purpose, are provided by Stratagene. Stratagene shall have
no liability for any direct, indirect, consequential, or incidental damages arising out of the use, the results of use, or the inability to use
this product.