Genetics Lab Section #1
In this lab investigation, an unknown sequence is given and by using specific techniques, the sequence can be identified and to which organism it belongs without laying a hand on a pipette. From the unknown sequence given, NCBI website is used to find the organism, which is a house mouse (Mus musculus). The sequence encodes the gene E2a. E2a is a transcription factor that regulates transcription. E2a is essential for the early B-lymphocytes development; they active the Ig gene (mimmunoglobulins) rearrangement and also play a role in class switch recombination. PCR will be used to amplify the given sequence. Once the sequence has been amplified to the desired amount, it will be double digested with EcoRI and BamHI, which leave a sticky ends; it will then be ligated to a plasmid Litmus 38 (plasmid also be double digested with EcoRI and Bam- HI).
The plasmid will then be transformed into E.coli where it will grow to make many colonies on a plated medium and then a mini-prep is used to recover the plasmids.
As geneticists continue on their path to understand how genes, RNAs, proteins, DNA, transcription factors, etc. function in each organism on Earth, many recombinant DNA techniques have been developed for these purposes. In this lab investigation, an unknown sequence is given and by using specific techniques, the sequence can be identified and to which organism it belongs. The purpose of this lab is to find what organism the unknown sequence belongs to and to amplify the sequence by doing PCR. In order to amplify the sequence, restriction enzymes, such as EcoRI and BamHI, and a plasmid is required. The restriction enzymes will cut but the plasmid, which will be ampicillin resistant (antibiotics), in two...