Manual: Universal Human Reference rna

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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.