Egfp Lab Report

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The Process of Ligation, Transformation, and Isolation of EGFP cDNA Into pET-41a(+) By Using Miniprep, Restriction Digest, PCR, and Bioinformatics To Ultimately Create Recombinant Expression Plasmids.
INTRODUCTION:
Scientists in this day and age are now able to change DNA and create recombinant DNA, which is a huge indication of great possibilities for the future of genetics. The goal of these lab experiments was to be able to create recombinant DNA with the insert of egfp gene. EGFP also known as Enhanced Green Fluorescent Protein was originally isolated from the jellyfish Aequorea Victoria and was introduced into the scientific field as a tool to signal gene expression. When EGFP is exposed to Ultraviolet Light, the protein glows a very bright green color. The egfp gene extracted from the jellyfish Aequorea Victoria was then modified to be able to be used in a lab setting in order to indicate gene
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Well #3 and #5 were very similar to each other with bandings at around 1.0-1.5kb, but also continual large lane banding. Well #4 of the non-recombinant has banding below 0.5kb. Well #6 was the Positive Control and showed one large lane band. Well #7 was the Negative Control that resulted in no bands because it was made up of H2O. The last well contained the DNA Ladder, which is used to measure the other band sizes.
Bioinformatics To Recreate An Online Recombinant Plasmid- The PCR product size was found to be 1183bp. The bold parts of the sequence at the start and end are the primers. A hypothetical Restriction Digest Design could use Acc65I and ArsI restriction enzymes to create cuts from the PCR product. The PCR product that was cut with only Acc65I would create 2 fragments of sizes 1-425bp and 426-1183bp. If the PCR product was cut with only ArsI, it would also create 2 fragments of sizes 1-723bp and 724-1183bp. The double digest would create 3 fragments of 1-425bp, 426-723bp, and

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