Page designed and maintained by Octavian Henegariu (Email: or ). WARNING: The information provided in these pages is copyrighted and is intended for individual use only. No parts of this work (text, tables or pictures) may be commercialized, published or otherwise reproduced without the written consent of the author. For a complete description of primers, PCR programs and a discussion of the PCR conditions please consult: Andrologia 26: 97-106 (1994) and Biotechniques 23: 504-511 (1997). Click here to get the Biotechniques paper in PDF format. PCR | dUTP label | FISH | FISH guide| CCK | Slide prep | CM-FISH | TM-FISH | µArrays| Home New links !! --> FISH guide | CCK procedure | Nucleotide labeling  Taq polymerase Different concentrations of a Taq polymerase were tested using primer mixture C (Fig. 32). The most efficient enzyme concentration seemed to be around 0.4µl or 2 Units/25µl reaction volume. Too much enzyme, possibly because of the high glycerol concentration in the stock solution, resulted in an unbalanced amplification of various loci and a slight increase in the background. too little enzyme resulted in the lack of some of the amplification products. Fig. 32. Amplification products of mixture C, using 0.5 Units/25µl, 1 Unit/25µl, 2Units/25µl, 4 Units/25µl and 8 Units/25µl reaction volume are shown. Arrows indicate the expected positions of the amplification products. The most appropriate enzyme concentration was between 1-2 Units/25µl. Five native Taq polymerases, from five different sources, were used to amplify multiplex mixture D in 1.6x PCR buffer using 2Units enzyme/25µl reaction (Fig. 33). In the same buffering conditions, all these enzyme performed similarly. Fig. 33. Multiplex PCR of mixture D in 1.6x PCR buffer using Taq polymerases from five sources. Lanes 1 to 5 indicate that all enzymes work similarly at the same concentration. Lane 4* (green) shows the products obtained when the enzyme from lane 4 was used in the buffer provided by the vendor. An unspecific product appeared, indicating that buffer composition influences the results.