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Rfp Lab Report

700 Words3 Pages

The presence of either a RFP or GFP gene in a plasmid DNA containing the pQE30 vector is determined using PCR and confirmed using Sanger DNA sequencing analysis. Two PCR reactions were performed; one using a primer for RTP and the other using a primer for GTP. Upon completion of 25 cycles of PCR, gel electrophoresis was performed to determine the size of PCR product and if amplification occurred. The reaction containing RF and RR primers showed amplification indicating that the DNA was encoded with the gene for RFP. With this knowledge, another sample was created for DNA sequencing using the Plasmid DNA and primers for RFP. Upon translating the gene sequence, presence of the RFP amino acid sequence confirmed the presence of the gene and successful PCR. Introduction The purpose of this lab was to use polymerase chain reaction (PCR) to amplify small amounts of DNA with two primers specific to two different genes: the red fluorescent protein (RFP) and green fluorescent protein (GFP). The DNA contains a specific sequence that is identified and annealed to by a specific primer. Only one of the two reactions should show amplification, identifying the presence of RFP or GFP. …show more content…

It involves annealing two complementary primers using hybridization to a single DNA strand and using DNA polymerase to extend the primer thus replicating the DNA. This cycle is repeated, in this experiment, 25 times between initial denaturation and final extension using a thermal cycler and as a result, the desired DNA sequence is amplified 2^25 fold. This revolutionary technique allows the study of structural and/or functional gene mutations, the ability to clone and manipulate desired genes and DNA analysis of archaeological

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