CulGenex™

Electrophoresis Buffer Powders

Cat. no. C4618 CulGenex™ TBE (Tris Borate EDTA), 10X, 1L Pouch 6 pouches/pack

INTENDED USE

Hardy Diagnostics CulGenex™ TBE Electrophoresis Buffer Powder is used for agarose gel electrophoresis in the analyses of nucleic acid products resulting from PCR amplification, purification protocols, or cloning experiments.

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

TBE Buffer is often used for polyacrylamide and agarose gel electrophoresis. It is particularly useful for the separation of smaller DNA fragments, such as the products of restriction enzyme digestion. It is ideally suited for DNA and RNA separation on longer run, higher voltage or amperage gels. TBE has a greater buffering capacity and will provide more sharply defined band resolutions than TAE Buffer. However, TBE may inhibit DNA ligase which may cause problems if subsequent DNA purification and ligation steps are intended.

Hardy Diagnostics CulGenex™ TBE (Tris Borate EDTA), 10X powder comes conveniently packaged in a 1 liter pouch and is ready to mix at the 10X concentration for easy dilution to a 1X working solution. It is available in packages of six.

FORMULA*

TBE (Tris Borate EDTA), 10X:
Gram weight per liter: 170.0gm/L
Tris-Base 0.8M
Boric Acid 0.89M
EDTA 0.02M

Final pH 8.3 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Store the sealed pouches containing dehydrated powder(s) at 2-30ºC. Dehydrated powder(s) can be very hygroscopic. Keep lid tightly sealed. Protect dehydrated powder from moisture and light. The dehydrated powder(s) should be discarded if it is not free-flowing or if the color has changed from its original white.

Store the prepared buffer at 2-30ºC.


PRECAUTIONS


METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend the contents of one pouch of powder (170.0gm/L for TBE Buffer) in molecular grade water (Cat. no. CG490 or CG490BX). QS to one liter and stir to mix thoroughly. This prepares a 10X solution of buffer.

2. Heat as necessary to dissolve completely.

3. Sterilize by autoclaving at 121ºC. for 15 minutes.

4. Cool to 45-50ºC.

PROCEDURE

Consult listed references for information on specific protocols.(1-5)

LIMITATIONS

Note: Failure to dilute 10X TBE to a 1X working solution will result in very slow migration of the samples and very high amperage (causing excessive heating of the gel).


MATERIALS REQUIRED BUT NOT PROVIDED

Agarose LE, Molecular Biology Grade (Cat. no. C8740 and C8741) powder, Agarose, Fine Resolution, PCR Grade (Cat. no. C8750 and C8751) powder, Agarose, Low Melt (Cat. no. C8760 and C8761) powder sold separately.

QUALITY CONTROL

Hardy Diagnostics tests CulGenex™ TBE (Tris Borate EDTA), 10X for nuclease activity, protease activity and pH.

User Quality Control


Physical Appearance

CulGenex™ TBE (Tris Borate EDTA), 10X powder should appear homogeneous, free-flowing, and white in color. The prepared buffer should appear clear, and colorless.

REFERENCES

1. Ausubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, K. Struhl, Editors. 2010. Current Protocols in Molecular Biology . John Wiley and Sons, Inc. Malden, MA.

2. Bullock, G.R. and Petrusz, P. 1989. Techniques in Immunocytochemistry, Volumes 1, 2, 3 and 4, Academic Press, London.

3. Cseke, L.J., P.B. Kaufman, G.K. Podila, and C.J. Tsai. 2004. Handbook of Molecular and Cellular Methods in Biology and Medicine . CRC Press.Taylor & Francis LLC. Boca Raton, FL.

4. Sambrook and Russell. 2001. Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press. Woodbury, New York.

5. Walker, J.M. 1984. Methods in Molecular Biology. The Humana Press Inc. Clifton, NJ.


111716vr