Essay On Agarose Gel

1205 Words3 Pages

In the preparation of agarose gel, agarose powder will be mixed with buffer. Agarose powders need to be weighed first before mixing it with buffer. When 1 gram agarose gel is added, it will be followed by 100 ml of buffer. The ratio is 1:100 for agarose powder and buffer respectively. Then, the solution of agarose powder and buffer is put in the microwave in order to melt the agarose until the solution become clear (Carson & Robertson, 2005).
Agarose gel is poured in the casting trays. Actually casting trays is available various in sizes and made of UV transparent plastic. Finally, the comb is used to make well in the gel where the dyed DNA will be loaded. According to Bio Rad (n.d.) comb is placed in slots on the casting tray. Usually, it is put in the slot before agarose gel (melted gel) is poured. Then, the comb is taken out after the gel solidifies. The comb will leave small hole that are called well that made from teeth of the comb.
TAE buffer (Tris-acetate-EDTA) 50X is use in this experiment of nucleic acids’ electrophoresis whether in agarose or polyacrylamide gels (Bisen, 2014). Usually for resolution of RNA and DNA fragment larger than 1500 bp TAE buffer is preferred, for genomic DNA and for large supercoiled DNA. In addition, TAE buffer contain low buffering capacity. Therefore, during prolonged electrophoresis, replacement (periodic) of the buffer is recommended. Furthermore, TBE buffer and also known as Tris-borate-EDTA 10X commonly used in polyacrylamide gel electrophoresis for the DNA and RNA (Kumar & Garg, 2005). TBE buffer is also used for agarose gels but it is rarely used for preparative gels for recovery of nucleic acids. TBE buffer will act as a strong inhibitor for nucleases. Double stranded linear nucleic a...

... middle of paper ...

...ample is occurred. According to Protocol: Preparation of agarose gel for DNA analysis (2013), the enzyme of DNase can also be release from our hand and this can infect our DNA by degradation by that enzyme. This glove also can be useful when deal with Ethidium bromide; since EtBr intercalated dye is very toxic and can cause infertility, we must do take care of our self when deal with this EtBr. After finishing this AGE Analysis, we must quickly remove the gloves and throw it in the sterilized bin.
Lastly, when the gel is viewed with an ultraviolet (UV) transilluminator, the high wavelength of UV light can hurt our body. So before UV light box is set upon, we must put on UV protecting face shield and make sure our body are secure from UV exposure. The exposure of the UV light also must be restricted because exposure over the time limit will cause the DNA to damage.

Open Document