1 introduction, 2 criterion tbe-urea gels, 1 gel composition – Bio-Rad Criterion™ TBE-Urea Precast Gels User Manual
Page 33: 2 gel selection guide, 3 tbe-urea buffers, 4 sample preparation, Denaturing nucleic acid page

Denaturing Nucleic
Acid PAGE
10
10.1 Introduction
Criterion
™
TBE-urea gels are used for separation of small RNA and single-stranded DNA (ssDNA)
fragments. Applications include oligonucleotide analysis, RNase protection assays, and northern
blotting.
10.2 Criterion TBE-Urea Gels
10.2.1 Gel Composition
Gel buffer
89 mM Tris, 89 mM boric acid, 2 mM EDTA, 7 M urea, pH 8.3
Crosslinker
3.3% C
Stacking gel
4% T, 3.3% C
Storage buffer
89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8.3
Shelf life
~8 weeks at 2–8°C; expiration date is printed on each cassette
10.2.2 Gel Selection Guide
Gel Percentage
Optimum Separation Range
5%
50–1,000 nt
10%
25–300 nt
15%
10–50 nt
10.3 TBE-Urea Buffers
See Appendix B for buffer formulations. Do not adjust pH unless directed to do so.
Running buffer (1x)
89 mM Tris, 89 mM boric acid, 2 mM EDTA
Dilute 100 ml 10x stock (catalog #161-0733) with 900 ml diH
2
O
Sample buffer (5x)
89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8.0, 12% Ficoll,
(catalog #161-0768)
0.01% bromophenol blue, 0.02% xylene cyanole FF, 7 M urea
10.4 Sample Preparation
Determine the desired ssDNA or RNA concentration for your sample based on the detection method
used. Dilute 4 parts sample with 1 part sample buffer.
Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com
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