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The serial transfer experiment with more than one sample


The 3SR method
Manfred Eigen Scientist
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There are other methods besides PCR, but PCR was the first method and it was Kary B Mullis who established it really, and got a Nobel Prize for it, although very important work in this field also was done in Gobind Khorana's laboratory. Now the other method... I perhaps should mention another one, which is called the 3SR method. I don't tell the abbreviation but I rather describe the method. You need three enzymes for the method, but that's not the 3SR, it does not refer to three enzymes. But what is it? It's first of all an isothermal method... you don't have to cyclise the temperature. Second, you start with the reverse transcriptase, that's the enzyme of the AIDS virus. That is able to transcribe the RNA back into DNA. You might remember that the dogma of molecular biology is that the information is in DNA, that's in a stable form there, that is transcribed into RNA where it's in a labile form and the RNA informs the protein factories, so that protein comes out of that DNA RNA protein.

But meanwhile one found an enzyme which can re-transcribe the RNA into DNA, and that's a whole class of viruses utilises this enzyme. They are called the retroviruses... utilises this enzyme to infect the cell, re-transcribe their RNA into DNA which then can be incorporated into the genome of the cell, though that's a very diabolic way of establishing the wrong genes in organisms. Yes, using this enzyme means that you start with an RNA molecule, you re-transcribe it into DNA, and then you take another enzyme, which is a so-called T7 polymerase, which means it is taken from a phage T7, and it is the polymerase which copies DNA and makes RNA. So you first re-transcribe the RNA into DNA, and now the polymerase comes and makes about one hundred RNA molecules from that DNA molecule. Now most of that RNA you decompose then by a nuclease, but before you decompose it you re-transcribe your amplified RNA into DNA and again each DNA makes one hundred RNA. You see while in the PCR reaction you make two, four, eight, sixteen, here you make 100 in the first step when the DNA is copied into RNA, in the next step you make 100 x 100, or perhaps a few less than that, and so you have a much steeper increase which for many of our methods turned out to be of advantage.

[Q] And the quality is the same?

Yes. Yes, the quality is... well, it's even easier to handle because you can do all your work at 40°C, whereas the other you have to heat up to nearly the boiling point, to above 90°C. And there are more, even other, amplification methods, I will not go into all the details, but the existence of these amplification methods was important for our evolutionary technology.

Nobel Prize winning German biophysical chemist, Manfred Eigen (1927-2019), was best known for his work on fast chemical reactions and his development of ways to accurately measure these reactions down to the nearest billionth of a second. He published over 100 papers with topics ranging from hydrogen bridges of nucleic acids to the storage of information in the central nervous system.

Listeners: Ruthild Winkler-Oswatitch

Ruthild Winkler-Oswatitsch is the eldest daughter of the Austrian physicist Klaus Osatitsch, an internationally renowned expert in gas dynamics, and his wife Hedwig Oswatitsch-Klabinus. She was born in the German university town of Göttingen where her father worked at the Kaiser Wilhelm Institute of Aerodynamics under Ludwig Prandtl. After World War II she was educated in Stockholm, Sweden, where her father was then a research scientist and lecturer at the Royal Institute of Technology.

In 1961 Ruthild Winkler-Oswatitsch enrolled in Chemistry at the Technical University of Vienna where she received her PhD in 1969 with a dissertation on "Fast complex reactions of alkali ions with biological membrane carriers". The experimental work for her thesis was carried out at the Max Planck Institute for Physical Chemistry in Göttingen under Manfred Eigen.

From 1971 to the present Ruthild Winkler-Oswatitsch has been working as a research scientist at the Max Planck Institute in Göttingen in the Department of Chemical Kinetics which is headed by Manfred Eigen. Her interest was first focused on an application of relaxation techniques to the study of fast biological reactions. Thereafter, she engaged in theoretical studies on molecular evolution and developed game models for representing the underlying chemical proceses. Together with Manfred Eigen she wrote the widely noted book, "Laws of the Game" (Alfred A. Knopf Inc. 1981 and Princeton University Press, 1993). Her more recent studies were concerned with comparative sequence analysis of nucleic acids in order to find out the age of the genetic code and the time course of the early evolution of life. For the last decade she has been successfully establishing industrial applications in the field of evolutionary biotechnology.

Tags: 3SR, self-sustained sequence replication, Polymerase chain reaction, PCR, isothermal method, RNA, DNA, AIDS virus, T7 polymerase, phage T7, retroviruses, reverse transcriptase, evolutionary technology, Nobel Prize, Kary Banks Mullis, Har Gobind Khorana

Duration: 4 minutes, 6 seconds

Date story recorded: July 1997

Date story went live: 24 January 2008