Week 7 DNA Electrophoresis Lab

Directions: The following story is about a crime solved through biotechnological techniques. The techniques used are DNA fingerprinting and polymerase chain reaction (PCR). DNA fingerprinting is a technique in which an individual’s DNA is analyzed to reveal the pattern of particular short nucleotide sequences. This pattern is claimed to be unique to the individual concerned and can thus be used for identification purposes. Polymerase chain reaction, on the other hand, is a technique used to replicate a DNA fragment so as to produce many copies of a particular DNA sequence. Why do you think there’s a need to generate several copies of the DNA? You will learn the answers to this as you go through the story. Read the story carefully and answer the questions that follow.   Questions: 1. How did the investigators conclude that the suspect was indeed the murderer?  2. What if a strand of hair was found in the truck bed instead of a seed pod, do you think it will still lead to the…

Directions: The following story is about a crime solved through biotechnological techniques. The techniques used are DNA fingerprinting and polymerase chain reaction (PCR). DNA fingerprinting is a technique in which an individual’s DNA is analyzed to reveal the pattern of particular short nucleotide sequences. This pattern is claimed to be unique to the individual concerned and can thus be used for identification purposes. Polymerase chain reaction, on the other hand, is a technique used to replicate a DNA fragment so as to produce many copies of a particular DNA sequence. Why do you think there’s a need to generate several copies of the DNA? You will learn the answers to this as you go through the story. Read the story carefully and answer the questions that follow.

DNA EXTRACTION Materials: knife                                                               1 cup of fruit                              Table salt                                                       clean piece of cloth or strainer for filtering resealable bag                                              dishwashing liquid 70% rubbing alcohol (chilled)                   shot glass (or any transparent and narrow container resembling a test tube)                                                                                                                           How to do the extraction: Before you begin, make sure you have chilled your alcohol in the freezer for at least 3 hours. Room temperature alcohol will not work nearly as well as cold alcohol. Place about 50 mL of alcohol in the freezer to chill it and take it out only when you’re going to need it. (The alcohol will not actually freeze.)   1. The first thing you will need is a sample. Since DNA is found in all living…

Can you please check my answer and make sure it is correct. Question: List the ingredients of master mix, and state the purpose of each ingredient.  Answer: Taq DNA polymerase This enzyme synthesizes the complementary strand of the DNA template after attaching to the primer. This means that it adds on free nucleotides to the existing strand, but helps speed up the covalent bonding between these newly added nucleotides. This enzyme is also thermally stable meaning that it can withstand the hot temperatures needed for PCR to occur. This hot temperature is needed for the denaturation step when the double stranded DNA has to be unwound and separated into two strands.  Individual building blocks of DNA (either free nucleotides A, T, C, and G or dNTP’s) These nucleotides are needed to build the complementary strand of DNA A special buffer to maintain the optimal pH, salts, and MgCl2 These buffers help maintain a good pH that doesn’t become too acidic or basic for Taq DNA polymerase to…

Can you please check my answer and make sure it is correct.   Question: Describe the two roles primers play in PCR.  Answer: Primers are short sequences of single stranded DNA that mark both the ends of the target sequence to be amplified during PCR. One primer attaches to the top strand at one end (forward primer) and the other one attaches to the bottom strand at the other end of the segment (reverse primer). They are made specifically to attach to these sections of DNA because the nucleotides of the primers are complementary to the target sequence. Other than marking the starting and end points for PCR, primers are also necessary for the process because without them, DNA polymerase wouldn’t be able to synthesize the complementary strand of DNA because it can only add nucleotides to an existing piece.

Please I want the answer of (B) and (C) Task 1. Your graduate advisor asks you to amplify the following sequence of DNA by PCR:   5’-ATACGCATTCGGACCAGGTCCTAA-3’   3’-TATGCGTAAGCCTGGTCCAGGATT-5’   a. To ensure that the entire sequence above is amplified, what 6-nucleotide DNA primers   should you add to your PCR mix?       You order the primers listed above, but instead receive the following set of primers:   5’-CGCATT-3’   5’-GGACCT-3’   b. What portion of the double stranded DNA molecule will be amplified after 10 rounds of   PCR?       Your labmate attempts to rescue your PCR reaction by providing you with the following   set of primers:   5’-ATACGC-3’   5’-TCCTAA-3’   c. What is the result of running the PCR reaction with your labmate’s primers? How many   double stranded molecules of DNA will result from 10 rounds of amplification?

Why PCR needs the following: DNA template Primers DNA polymerase enzymes dNTPs Buffer Mg2+

Can you please check my answer and make sure it is correct.    Question: How can DNA evidence be used to convict or exonerate a defendant? Why is DNA evidence so powerful?    Answer: DNA evidence can be used to perform DNA profiling to determine the genotype of the specific DNA sample. With just a small amount of DNA, PCR can produce billions of copies of that specific segment. The segments that are used are from non-coding regions that contain STR’s or short tandem repeats. These very short DNA sequences are repeated and are specific to individuals because we inherit them from our mother and father. Gel electrophoresis separates the PCR products based on their size and each band is compared to the allele ladder. This process helps to identify the alleles present in the original samples. DNA profiling is performed at many loci to be able to tell the genetic difference between different individuals with a lot of certainty. The DNA from the different suspects is compared to the allele…

The following statements are either accurate as written or contain some errors. You need to rewrite each as an accurate statement. Make sure that you do not change the material the sentence is about. Highlight or underline any changes that you make to statements. Thank you! 1. Base Excision repair happens during replication and is when an incorrect nucleotide is incorporated into the DNA. During this type of repair the damaged or incorrect base is removed and fixed via Nick translation.  2. There are four levels of polypeptide structure: primary, secondary, tertiary, and quaternary. The secondary structure describes the linear sequence of the proteins in the polypeptide.  3. The majority of proteins fold spontaneously when they come out of the ribosome after translation. The reaming proteins need help from chaperones OR chaperonins to fold properly.

In a PCR-based crime scene investigation, similar to the one presented in the lab module with Brother Y and Brother X, there is a sample of DNA from a crime scene that is likely to belong to the guilty party. Based on the gel photo below, which shows the results of an electrophoresis gel following PCR amplification at one locus of 5 DNA samples - one crime scene sample and 4 suspects - which suspects can be excluded from this investigation? [Keep in mind that it is not possible for a  heterozygous person to leave only one allele at a crime scene. If any one allele does not match, then that suspect is eliminated.] Choose all that apply.

Can you please check my answer and see if it is correct.    Question: In addition to master mix, what else must be added to each PCR tube? Why?    Answer: In addition to master mix, the primers must be added to the PCR tubes. Each tube is also reserved for one individual’s specific section of DNA. So not only do primers needed to be added that are made specifically for the person’s DNA, but the DNA itself from the suspect needs to be added to the PCR tubes (DNA template strand). We also must have a control which contains sterile water, and one PCR tube reserved for DNA from the crime scene.

5) Answers to the following questions. A. You are given a tube containing 275 ng of purified PCR product (DNA) that is 1262 bp long. How many picomoles of PCR product are in the tube? The average weight of a deoxynucleotide monophosphate is 328 g/mol. B. You digest a 5 kb plasmid with two enzymes that yield fragments of 4 kb and 1 kb, and resolve the digest on the gel. What would you predict to see in terms of the intensities of the bands?

BIOTECHNOLGY Date: Name: Instructor: Section/Group:. POST-LAB QUESTIONS 1. In one or two sentences, summarize the technique of gel electrophoresis. 2. How does the process of gel electrophoresis separate DNA fragments? 3. Why is the fact that DNA has a negative charge so important in the gel electrophoresis process? Biotechnology 165

Your next task is to study Ebola virus (EboV) sequences of different strains. You have received sequences of different Ebola virus strains collected during the Ebola outbreak of 2013 – 2016.  EboV from Guinea pig Reference DNA Sample  CTA TGC AAG CAG TTA mRNA           Protein

Paternity Test: Alanna claims John is the father of Timmy, but Leo thinks that he is the father. All of their DNA samples were taken and cut with the same restriction enzyme and run through a gel. 4. Who is the father of Timmy? 5. How many of Timmy's fragments are the same size as his mother's fragments? 6. Whatactually causes the DNA pieces to move through the gel? 7. Describe the difference you see in the gel between John's DNA and Leo's DNA. Alanna's DNA Timmy's DNA John's DNA Leo's DNA

Which of the following are necessary for Sanger sequencing? Select all correct answers. Group of answer choices ddNTPs an oligonucleotide primer DNA polymerase dNTPs DNA ligase a restriction enzyme

DNA is visualized during agarose gel electrophoresis by ______________ . the fact that DNA fluoresces when illuminated with UV light the binding of a fluorescent dye that is easily detectable using radioactive antibodies that specifically bind to DNA the fact that DNA is blue and can be seen when millions of copies are present in a band

what is the |usage of DNA separation in gel electrophoresis please write the resources under the answer

You need to complete the following steps for an experiment. Explain whether an acrylamide gel or an agarose gel is more appropriate to use for the following experiments. Purifying tRNA from total RNA Isolating a vector insert for cloning Analyzing PCR products

DNA Sample : Blood Stain collected from handle of knife blood Bob Sue John Lisa stain 1. What do you notice about the black lines? 2. How do we read the gel electrophoresis? 3. Which sample are we comparing Bob, Sue, John, and Lisa's DNA to? 4. How do they compare?

DNA extraction, PCR, gel electrophoresis, and DNA sequencing of PCR products. Give a description of these methods and the materials required to perform each of these molecular techniques

I need help defining what is DNA Chromatography.

You are studying the DNA binding protein CLAMP and you want to determine its binding affinity for different DNA constructs, what technique should you use? ELISA PCR Chromatography Western Blotting Fluorescence polarization

DNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at FC or at room temperature. Longer spins make it difficult to resuspend cells. 2 Resuspend pellet in 100pul GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200ul NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150ul potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…

The following gel shows the results of a crime scene investigation. Lane 1 shows a DNA sample that was obtained from the scene of a crime (there was evidence of the criminal cutting himself, so it's the criminal's blood sample that produced the DNA bands in lane 1). Lanes 2-7 shows a DNA sample from six potential suspects held in connection with the crime. Arrange the bands labeled A, B, C, D from largest to smallest. 1 2 3 4 5 6 7 OC, A, D, B A, B, C, D OB, D, A, C O D, A, C, B

Answer the following questions related to PCR Bioinformatics for DNA Extraction using  Why is it necessary to chelate the metal ions from the solution during the boiling/lysis step at 100 degrees celsius? What would happen if you did not use a chelating agent such as the InstaGene matrix? What is needed from the cells for PCR? What structures must be broken to release the DNA from a cell?

Describe and contrast the common steps of DNA replication in vivo and the PCR reaction in vitro? In simple terms so, that I can understand. Thank you

Figure 9-22 shows the first steps in the process of making a DNA microarray, or DNA chip, using photolithography. Describe the remaining steps needed to obtain the desired sequences (a different fournucleotide sequence on each of the four spots) shown in the first panel of the figure. After each step, give the resulting nucleotide sequence attached at each spot.

schematic diagram for the procedure in doing DNA extraction, isolation and quantification

From your knowledge about DNA microarray, answer the following: Why RT-PCR is important in the sample preparation to perform expression microarray experiment?(

What enzyme proofreads the DNA molecule?   Group of answer choices DNA gyrase DNA polymerase I DNA helicase DNA polymerase III

Which of the following components are required for the polymerase chain reaction (PCR)? Select all correct answers. Group of answer choices dNTPs a restriction enzyme DNA polymerase DNA ligase ddNTPs oligonucleotide primers

For what purposes is DNA extraction done? (give at least 3 purposes for which you may need to extract DNA)