Prior to the preparation of the gel, the casting tray was inserted into the electrophoresis chamber. The tray was put into the chamber with the orange gaskets against the side walls. The comb was placed into the first set of slots. The slots are located at the top of the tray. During these procedures, the researcher made sure that one can clearly read 1.5 on the left side of the comb when placed properly in the tray. 40 ml was the volume of agarose gel solution needed for the purposes of this procedure.
The right quantity of agarose to make a 0.8% solution was measured out and placed in a 125ml Erlenmeyer flask. During the measurement, it was determined that agarose gels are cast as a percent weight per volume solutions (%w/v). For example, 100 mls of a 2% (w/v) solution is 2g/100ml. An appropriate amount of 50X TAE was added into the flask so that the final concentration was 1X. 35ml dH2O was then added into the flask and the contents swirled gently. The flask was then placed in the microwave and heated for 1 minute. After heating, the flask was removed, swirled gently, and put back into the microwave for an additional 30 seconds.
After heating, the flask was allowed to cool for 2 minutes before its contents were poured into a 50ml graduated cylinder. The volume of the flask was brought to 40mls through the addition of dH2O. The gel solution was returned back into the flask. 1Âµl of ethidium bromide was added into the combination. The researcher took caution when handling ethidium bromide as it is a hazardous substance. For example, the substance is a known mutagen and a suspected carcinogen. As a result of this, gloves were worn when handling it. The gel solution was then poured into the casting tray and allowed to sit for 15 minutes for it to solidify. The gel turned opaque with a slight blue hue when it was ready.
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