Comments
For the separation of oligonucleotides with a small number of bases, acrylamide gels with a small pore size are used. The pore size is determined by the total concentration of acrylamide and the ratio of acrylamide : bisacrylamide. For the sequencing of nucleic acids a ratio of acrylamide : bisacrylamide of 19 : 1 is used. Caution: Acrylamide is a potent neurotoxin and absorbed through the skin. The effects of acrylamide are cumulative. Although polyacrylamide is suspected to be nontoxic, it should be handled with great care because of the possibility that it might contain small quantities of unpolymerized acrylamide.
Literature
(1) Peacock, A.C. & Dingman, C.W. (1967) Biochemistry 7, 668-674. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. (2) Ambrose, B.J.B. & Pless, R.C. (1987) Methods Enzymol. 152, 522-538. DNA sequencing: Chemical methods. (3) Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. (4) Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. & Struhl, K. (eds.) (1995) Current Protocols in Molecular Biology. Greene Publishing & Wiley-Interscience, New York.
