Kuntaman, - and Indrayana N.S., - (2002) Propagasi Lambda Faga Menggunakan Teknik Transformasi-Transfeksi Sebagai Langka Awal Penelitian Biologimolekuler. Other thesis, UNIVERSITAS AIRLANGGA.

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Abstract

Gel electrophoresis is an essential step on molecular biology research. The one important step is how to measure the DNA fragment size, that need the DNA Marker. The selected substance that is usually used as a source for making of the DNA Marker is Lambda phage DNA. Commercially product of it had been sold elsewhere, and is made by cutting Lambda phage DNA using HindIII enzyme. The aim of this research is how to propagate Lambda phage DNA by using method of transformation-transfection. This research was conducted in Laboratory of Tropical Disease Center (TDC) Airlangga University. Lambda phage DNA was get from Sigma and Escherichia coli strain LE-392 was used as a host. The competent cell of Escherichia coli was prepared by method of Calcium Chloride treatment. Heat shock was done by heating 42 C for two minutes. Prior for transfection, Escherichia coli LE392 was cultured overnight in LB broth with addition of Maltose 0.2% and MgSO4 1 mM. Culture of phage infected Escherichia coli was conducted on top agar. Isolation of phage DNA was performed by scrabing of top agar by CMS solution and DNA isolation by using STE and PEG-8000 solution. The result shows that after overnight incubation, was identified as much as 20 plaque per plate or 400 phage infected Escherichia coli per mililiter solution. After sub-culture of phage infected Escherichia coli with addition of maltose induced Escherichia coli LE392, was get abundant of plaque on the plate culture. The isolation of genome of phage from top agar shows a band of DNA in the same size with the original Lambda phage DNA size.

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