QBM Exam 3 Questions And Answers With Complete Solutions 2024

QBM Exam 3 Questions And Answers With Complete Solutions 2024 Chapter 13 - DNA Purification, Agarose Gel Electrophoresis, and DNA Visualization - answer The genetic information of all living organisms is contained within the sequence of ____ ____ = Examples include: purines and pyrimidines = For most organisms, the genetic information is contained in nucleic acids of the deoxyribonucleic acid (DNA), or ribonucleic acid (RNA) in certain viruses - answernucleic acids ______ is a complete set of nucleic acids containing the genes of an organism. = Example: There are 5 million base pairs in E.Coli meaning 4300 genes. = There are 3 million base pairs in Human genome ( 21,000 genes) - answergenomes Since an individual gene constitutes only a small fraction of the GENOMES, the study of a gene often starts with the manipulating the gene to ______ it. = Another methods is to monitor the expression of it. What we do it we place the gene of interest into a _______ = which is a extrachromosomal DNA. Wait for it to be expressed to examine the gene of interest. - answerThe study of a gene often starts with manipulations designed to PURIFY the gene or to monitor the expression. Place purify gene into Plasmid! = Plasmid: extrachromosomal DNA Will know all of this by end of chapter! How do you purified an entire genome and NOT just one gene alone? (7 ways) - answer= DNA manipulation (cloning) = Gene libraries = PCR = Hybridizations - Dot Blot - Southern Blot = RFLP, VNTR = RAPD = Sequencing Unlike proteins, DNA can survive harsh condition, buffers, and reagent. - One of the main differences is that DNA PURIFICATION can be done using _____ solvents/reagents. = Examples: NaCl Ethanol Phenol Chloroform Centrifuge - answerORGANIC REAGENTS The overview of DNA purification process is. 1. The lysis and ____ of cells 2. Removal of _______/ purifying the DNA using reagents. = In most protocols, the enzyme used in degrading DNA is _______ ___. = This enzyme can also degrade proteins as well and is active in SDS, EDTA, UREA, and wide range of pH. - answer1. Lysis / disruption of cell 2. Removal of contaminate using reagents/ organic solvents will work. = Enzymes used for purification is usually Proteinase K. There are many protocols that can be used to purified DNA. Only focus on 1. Organic solvents (Phenol-Chloroform Extraction) 2. Alkaline Lysis = Miniprep 3. Silica-based methods - selective adsorption = Know the overview of these if you must 1. Crude lysate 2. Salting-out 3. Cesium chloride gradients 4. Anion exchange - answer 1. Crude Lysate = This method is when you use ____ on the cells and then add the enzyme ____ ____. = This is not a good method because DNA will still be somewhat dirty. = This method allow ____ steps. 2. Salting-Out = This method is when you use _____ ______ to salt out the DNA, and then centrifuge it and add alcohol to precipitation the DNA. 3. Cesium Chloride Gradients = Not really mentioned in the book, but it is very difficult, time consuming, and it require the use of _____ reagents. 4. Anion exchange = The _____ of the DNA will binds to the + charged beads. Then alcohol is added to precipitated it. - answer1. Crude Lysate: Heat and then Proteinase K = Allow downstream steps. 2. Salting Out: Potassium Acetate, centrifuge, Alcohol 3. Cesium Chloride Gradients: Toxic Reagents 4. Anions Exchange: DNA negative charged backbone to + charge of beads, alcohol to precipitated. Out of Crude Lysate, Salting-Out, Cesium Chloride Gradients, and Anion exchange. Which two use alcohol for precipitation? - answerSalting Out and Anion Exchange √√√ 1. Organic Solvents = Phenol- Chloroform Extraction = In this method uses _____, ______, and ____ _____ in a ___:___:___ ratios. = The Phenol is aqueous/organic which will contain ___ and ___. = The Chloroform is aqueous/organic will contain ____ and cellular debris (scattered fragments). = The process of Phenol-Chloroform is very tedious, produces hazardous ____, contain residual organic solvents that can inhibit the downstream process of _____. - answerPhenol Chloroform isoamyl alcohol Ratio: 25:24:1 = Phenol = Upper = aqueous = DNA and RNA = Chloroform = Lower = Organic = Proteins and cellular debris. = Produce hazardous waste, residual organic solvents, and inhibit PCR. PCR= Polymerase chain reaction (PCR) is a technique used to amplify a single copy or a few copies of a piece of DNA. √√√ 2. Alkaline Lysis = This method uses _____ to break up phospholipid bilayer. = And uses ______ to dissolve structural proteins.