The DNA Interpretation – Restriction Enzymes and Gel Electrophoresis BIO SCI 150

The DNA Interpretation – Restriction Enzymes and Gel Electrophoresis
BIO SCI 150
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I) Introduction
The deoxyribonucleic acid (DNA) is a molecule and nucleic acid, which stores and passes the biological information and is present in all living cells, the DNA carries the genetic information and encodes all proteins. The DNA consists of nucleotides molecules where the sugar-phosphate strands intertwine and form a double helix. Plasmids are circular mini chromosomes containing the protein coding sequence, and they may one contain one or multiple genes .The restriction endonuclease (enzymes) come from bacteria and they cut the DNA strands at specific sequences or recognition sites, and the enzymes recognize the specific sequences. The restriction endonuclease targets are derived from Escherichia coli and break the DNA molecule into fragments. I hypothesized that the nucleotide sequences for the two plastids would differ.
II) Methods:
In restriction digestion EcoRI is one of the restriction enzymes that cuts DNA at specific sequences, the 10 µl of pEC16S and 10 µl of pVH16S were the plasmids used in the experiment, while the 10 µl of EcoRI was the restriction enzyme and 10 µl of Pstl is a restriction enzyme that serves as a control.
Making the gel
The 1% agarose gel is used where 0.4g of the gel is put on an Erlenmeyer flask,
40 ml of TAE is added, the mixture is put in a microwave for 2 minutes
The mixture will boil and process is repeated until the agarose is resolved
There is cooling up to 60 o C, and after this the 4 µl of ethidium bromide (EtBr) is added
The mixture is put in a gel plate with the comb still present, and after 10-15 minutes the gel ...