Molecular Biology

To meet highest demands for quality, we offer a wide range of BioUltra Reagents for molecular biology.

Introduction

It has been well recognized that the use of basic reagents of highest purity is a definite prerequisite for successful work in molecular biology. Therefore many authors of laboratory protocols recommend purification of commercially available basic reagents in order to eliminate interfering traces of chemical impurities. Well known examples are the denaturation reagents phenol (1), formamide (2) and DMSO (2). Additional problems may be caused by contamination of the basic chemicals with traces of enzyme impurities such as DNases, RNases, phosphatases and proteases. Hence time consuming pretreatment of the basic reagents is necessary. To meet the increasing demand for such high purity reagents used in gene technology experiments, Fluka has recently introduced a specific BioUltra quality grade: BioUltra for molecular biology. Products of this group are free of both interfering chemical traces and the above-mentioned enzyme impurities.

Use of this New Quality Grade

Products belonging to the quality grade BioUltra for molecular biology are wellsuited for all commonly used basic experimental methods in the field of molecular cloning. This includes methods and techniques such as:

• DNA purification (plasmids, bacteriophages etc.)
• RNA purification
• precipitation of nucleic acids
• density gradient centrifugation
• electrophoresis
• transformation/transfection
• screening by hybridization
• Northern and Southern hybridization
• sequencing
• PCR amplification
• preparation of storage solutions
• (for enzymes, vectors etc.)
• preparation of enzyme buffers

Example: RNA Purification

The most important protocols for RNA isolations are based on the use of guanidine salts as deproteinization agents (2,3,4). Chomczynski and Sacchi (5) developed a single-step procedure which has become very popular since its publication in 1987. This and related protocols use guanidine thiocyanate (50990 (BioChemika) or 50980 (BioUltra for molecular biology)) as denaturants. Besides guanidine thiocyanate, Fluka has recently introduced all basic chemicals for RNA purification in the new quality BioUltra for molecular biology. Moreover, all the reagents used in the protocol of Chomczynski and Sacchi have been combined in a kit (Fluka 83913) and are offered together with a step-by-step protocol.

References

1. D.M. Wallace, Meth. Enzymol. 152, 33 (1987)
2. T. Maniatis et al., in Molecular Cloning. A. Laboratory Manual. CSH Lab. N.Y. (1989)
3. R.A. Cox, Meths. Enzymol. 12, part B (L. Grossmann, K. Moldave, eds.), 120 (1968)
4. J.M. Chirgwin et al., Biochemistry 18, 5294 (1979)
5. P. Chomczynski and N. Sacchi, Anal. Biochem. 162, 156 (1887)