BIO 111 Lab Final – TAMU (2023/2024) (Complete and Accurate)

BIO 111 Lab Final – TAMU (2023/2024) (Complete and Accurate) coding region comprised of genes that encode proteins (instructions that determine traits) and makes up 1-2% of human DNA. non-coding gene makes up 98-99% of human DNA, contains STRs and is responsible for regulatory functions such as gene transcription short tandem repeats (STRs) unique repeating patterns of the same nucleotide sequence. They can be used to differentiate one person from another. DNA Profile specific pattern of DNA attributes that is obtained in the lab and can be used to identify an individual. DNA profiling is the process of determining an individual's DNA characteristics (profile) from a sample of bodily tissue polymerase chain reaction (PCR) Purpose is to amplify DNA. In forensics, this can be used to obtain more DNA from a sample that would otherwise be too small to analyze What's inside master mix? 3 pairs of primers, loading dye, nucleotides (dNTPs), Taq polymerase, mix buffer, and water. DNA sample is added to master mix. Components of PCR Template, primers, taq polymerase, nucleotides, buffer template in PCR the DNA from the sample specimen serves as the template for replication primers in PCR short stretches of DNA that initiate the PCR reaction. Required for annealing and Taq binding at specific sites. Base sequences are complementary to the ends of the template DNA taq polymerase in PCR adds dNTPs to the template strand • Reads the original DNA sequence and creates a complimentary copy by adding in the new DNA bases nucleotides in PCR DNA bases (A, C, G, and T) serve as the building blocks of DNA and are used to assemble new strands of DNA buffer in PCR enables the reaction to take place by ensuring the right conditions are met (controls pH) What does Chelex do? The chelex resin chelates (absorbs) ions that inhibit function of the Taq polymerase enzyme what does the thermocycler do? Thermocyclers are instruments used to amplify DNA and RNA samples by the polymerase chain reaction. The thermocycler raises and lowers the temperature of the samples in a holding block in discrete, pre- programmed steps, allowing for denaturation and re- annealing of samples with various reagents. 3 steps of PCR cycle denaturation, annealing, extension Denaturation When the double stranded template DNA is heated to separate it into two single strands annealing when the temperature is lowered to enable the DNA primers to attach to the template strands extension when the temperature is raised and the Taq polymerase enzyme synthesizes new DNA strands by adding nucleotides to the template strands. purpose of lysing the DNA Lysis destroys the cell membrane and releases the contents of the cell into the solution what organism was taq polymerase isolated from? Thermus aquaticus: thermophilic bacterium (heat tolerant) that live in hot springs what is multiplex PCR? Amplification of multiple targets into a single PCR experiment by utilizing multiple primer pairs in a single reaction mixture Gel Electrophoresis Steps 1) Make the gel 2) Set up gel apparatus 3) Load DNA sample into gel 4) Hook up electric current and run gel 5) Stain