NA 100 R. Multi-functional electrophoresis device

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NA 100 R Multi-functional electrophoresis device No need for UV transilluminator and darkroom You can see DNA bands after 2 or 3 minutes of electrophoresis You can check

1 NA 100 R Multi-functional electrophoresis device No need for UV transilluminator and darkroom You can see DNA bands after 2 or 3 minutes of electrophoresis You can check 80 PCR products at a time. No need for exclusive camera for gel photo You can change the direction of electrophoresis. You can check and purify 20 PCR products at a time.

2 Characteristics of NA 100 R Bands of 80 PCR products confirmed at once and taken by a usual digital camera in NA 100 R

3 Sequencing data after purifying PCR product with NA 100 Molecular size marker, 1 bp ladder, from cloning the DNA purified by NA 100

Polarity converter: - Electrophoresis is in normal direction when the left side is pressed.

4 Components of NA 100 R 01. NA 100 R, multi-functional electrophoresis device, is composed of electrophoresis tank, UV transilluminator, DC converter, and polarity converter. Polarity converter: - Electrophoresis is in normal direction when the left side is pressed. - The direction is converted when the right side is pressed. UV: UV lamp S/W Life span of 6W UV lamp made in Japan is 3,000 hours. FN: Fan S/W for prevention of accumulating minute heat from UV lamp EP: S/W for electrophoresis 02. Agarose gel caster including the specially designed gel tray and combs for loading and collecting well

Micro-switch(OMRON, made in JAPAN) for safety, which is operated by electrophoresis tank cover or darkroom hood Manual of NA 100 R A. Making the agarose gel 01.

5 03. Transparent UV shield and electrophoresis tank cover that are specially designed to have dark room effect and to prevent being fogged up 04. Specially designed darkroom hood and filter for taking pictures with a usual digital camera 05. Micro-switch(OMRON, made in JAPAN) for safety, which is operated by electrophoresis tank cover or darkroom hood Manual of NA 100 R A. Making the agarose gel 01. Melt the agarose in microwave and then move the agarose gel to 50 or 60 oven more than one hour to prevent deformation of the gel tray by heat and to remove micro-air bubbles that are formed in process of melting the agarose. * The gel tray is highly UV transparent. But, it is easily deformed by heat(80 ), ethanol, xylene, or other organic solvents. 02. Assemble the gel tray and the gel caster. * If the combs are assembled with the gel tray and the gel caster at this step, there is high possibility of penetration at the bottom of well in the agarose gel. * In order to make the collecting wells, slightly push together the loading and collecting combs leftward or rightward after assembling the combs to make vertically parallel wells. 03. Pour the agarose gel into the gel tray. 04. Put the combs on the gel caster before the agarose gel becomes solid. 05. After solidifying the agarose gel, gently remove the combs from the gel

6 caster in perpendicular direction. 06. Remove the scraps of the agarose gel sticking to the bottom side of the gel tray. 07. Keep the gel tray with the agarose gel in the container of electrophoresis buffer mixed with Ethidium bromide. * When the agarose gel is separated from its tray, the gel will be easily broken, especially in the gel with collecting well. So, do not disassemble the agarose gel and its tray. 08. Before using the agarose gel, move it to another container of EtBr-free electrophoresis buffer. Then, remove the scraps of the agarose gel remaining in loading or collecting wells with the syringe. B. Electrophoresis 01. Check the polarity converting switch. Left side of S/W must be pressed. 02. Put the gel tray with the agarose gel on the dark blue part of the electrophoresis tank. 03. Fill the electrophoresis tank with electrophoresis buffer. 04. Load samples in the loading well. 05. Put the electrophoresis tank cover assembled with transparent UV shield on the electrophoresis tank. 06. Check the microswitch. You can hear sound, 'tick', when the microswitch is pressed by the electrophoresis tank cover. 07. Turn on the electrophoresis S/W(EP). 08. After 2 minutes, turn on the UV lamp and check the DNA bands. After checking the DNA bands, turn off the UV lamp. 09. Because migration velocity of DNA is influenced by the size of DNA and concentration of agarose gel, check the DNA bands after 5minutes or 10 minutes. * When the UV lamp is turned on for more than 2 or 3 minutes, turn on the fan S/W(FN) to prevent accumulation of the heat from UV lamp. 10. In order to take picture of DNA bands, replace the electrophoresis tank cover with the darkroom hood and press lightly darkroom hood for operation of microswitch(see Fig. B-1). * The flash of your digital camera must be off. * When the DNA bands are blurred, check the filter fogged up by vapors. If the filter of the darkroom hood is fogged up by vapors, wipe the

filter with the lens towel or soft paper such as Kimwipes R paper. And then take picture again. * Digital camera has various different focusing system according to its manufacturer.

pictures using the zoom lens of your digital camera(see Fig. B-2). Fig. B-1 Fig. B-2 C. Purification of DNA by NA 100 R 01.

7 filter with the lens towel or soft paper such as Kimwipes R paper. And then take picture again. * Digital camera has various different focusing system according to its manufacturer. If you have these difficulties in your digital camera, first, put the electrophoresis tank cover without UV shield on the electrophoresis tank and, second, put the darkroom hood, and then, take pictures using the zoom lens of your digital camera(see Fig. B-2). Fig. B-1 Fig. B-2 C. Purification of DNA by NA 100 R 01. Preparation of the agarose gel: * If the gel has been soaked in TAE, TBE, or TE buffer mixed with EtBr, empty out the loading and collecting wells. You can use a pipette to empty the loading or collecting wells, but much patience needed. Another better method is to use filter paper or kitchen paper towel folded fitly in long and narrow shape on the horizontal lane of the loading and collecting wells. The buffer of the loading and collecting wells will be easily removed. 02. Follow the step 01 and 02 of electrophoresis. 03. Fill the electrophoresis tank with electrophoresis buffer, 0.1X~1X TE, TAE or TBE according to your preference. Buffer should not be overflowed the upper surface of the agarose gel. Do not overfill! See the figure!

Medical Genetics and Diagnosis Lab #3. Gel electrophoresis

Medical Genetics and Diagnosis Lab #3 Gel electrophoresis Background Information Gel electrophoresis is the standard lab procedure for separating DNA by size (e.g. length in base pairs) for visualization