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Human Genome Lab Report

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Summary: The Sequence of the Human Genome
The Human Genome Project had been initiated by the National Institutes of Health and the U.S. Department of Energy with the goal of sequencing the entire human genome in 15 years. In order to sequence the human DNA, the DNA was obtained from five individuals, and plasmid clones were made using their DNA. The euchromatin of the human genome was sequenced using the whole-genome shotgun sequencing method. This method used mate pairs, which are paired-end sequences from subclone libraries. After using the whole-genome shotgun method, the sequenced DNA had to be assembled. Two assembly strategies were used- the whole-genome assembly and the regional chromosome assembly. The whole-genome assembly combines …show more content…

coli strain W1485, which was one of the materials used in this experiment. Many techniques were used such as DNA isolation, transformation, and restriction enzyme digestion. Restriction enzyme digestion by EcoRI gave a linear fragment on the electrophoresis gel because the plasmid had only one restriction enzyme sequence site. The generated size of the plasmid was the same as the predicted size.
The tetracycline-resistant gene of the R6-5 DNA may be separated from its site of replication by the EcoRI restriction site which can explain why tetracycline-resistant pSC101 plasmid was generated from R6-5 that underwent shearing, but no plasmids with tetracycline resistance were generated when R6-5 had been cleaved by EcoRI. Studies showed that R6-5 fragments that had been cut by EcoRI may be ligated in E.coli cells. These reassociated fragments can be functional plasmids if they are able to replicate, carry the antibiotic resistant gene, and able to form the circular shape of a plasmid.
The experiment was able to generate a mixture of plasmids pSC101 and pSC102. Because two plasmids were generated from R6-5 and were able to survive together in one host cell, it was assumed that R6-5 had at least two sites of

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