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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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The purpose of this lab was to perform a procedure known as genetic transformation which allowed us to genetically engineer E. Coli to be ampicillin resistance. Before the lab we expected that lysogeny broth and minus DNA will have growth but no glow. The lysogeny broth, ampicillin, and

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On the second day after these colonies were grown, the lab begun by labeling two test tubes -DNA and +DNA, adding the transformation solution to to each, and placing it on ice. Then the procedure called for the use of a sterile loop to pick up a colony of E coli and add it to each test tube, then after E coli was in both test tubes, the pGLO DNA was placed in ONLY the +DNA test tube. After 10 minutes on ice, and after the 4 plates are labeled each to LB -DNA, LB/amp -DNA, LB/amp +DNA, and LB/amp/ara +DNA respectively, the test tubes are heat shocked to loosen the DNA cell membrane to allow pGLO to slip in and code within the DNA. After more time in the ice bath, the tubes are taken out and set at room temperature with the LB broth placed in both test tubes. Then the dishes LB/amp +DNA and LB/amp/ara +DNA were spread with the +DNA solution, and the dishes LB -DNA, and LB/amp -DNA were spread with the -DNA solution.

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In 1984 the US Department of Energy, National Institutes of Health, and International groups held meetings to discuss studying the human genome. In 1990 the start of the Human Genome Project begins with the publication by the US Department of Energy and National Institute of Energy discussing the first five year plan and projecting it to be a 15 year project. Just five years in scientist discover for the first time that random “shotgun” sequencing can be applied to the whole genome with speed and accuracy. Shotgun sequencing is better known as shotgun cloning and it is used in complex long strands of the human genome. In 1996 the leaders of The Human Genome project decided that all human genomic sequence information should be free and available in a public domain within 24 hours of generation.

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