University of the West Indies at St. Augustine - BIOC 2169dna lab write up

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NAME: Latiyah Timothy LAB PARTNER: Jasiel Mohammed ID# 816012983 DATE: 28th January 2020 Course Code: Bioc 2169: Practical Skills in Biochemistry II Title of Lab: DNA extraction of genomic DNA from... fungi, plant, animal tissues using the CTAB method. Aim: 1. To extract DNA from a plant, fungi and animal tissues using CTAB method 2. To quantify DNA using spectrophotometric method 3. To determine the purity of the extracted DNA 4. To calculate the concentration of extracted DNA 5. To visualise extracted DNA using agarose gel electrophoresisTheory: Deoxyribonucleic acid or DNA is a molecule that stores genetic information. Its structure consists of molecules called nucleotides. The nucleotides, each contain a nitrogenous base a sugar group in this case deoxyribose and a phosphate group. There are four types of bases found in DNA, they are Thymine, Adenine, Cytosine and Guanine. These bases are attached together on a strand via phosphodiester bonds. The structure of DNA is that of a double helix. There are two single strands, running antiparallel to one another are joined together by the pairing of the bases (Adenine to Thymine and Guanine to Cytosine) via hydrogen bonding. In this lab, the DNA isolated is that from an animal, therefore, the DNA extracted is that of a right-handed double helix.[ CITATION Nel17 \l 1033 ] In this lab, DNA is extracted using the CTAB method. Cetyltrimethyl ammonium bromide (CTAB) is a detergent used to break open the cell walls and solubilise its contents. Then the cell membrane is disrupted to release the DNA into the extraction buffer. The extraction buffer contains EDTA and SDS (sodium dodecyl sulphate). SDS is a detergent used to inhibit the nucleases as well as separate the DNA from proteins. EDTA also inhibits the nucleases by chelating the magnesium ions that act as [Show More]

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