This is for an a 3 unknown I am doing in Microbiology. And I don't quite...
This is for an a 3 unknown I am doing in Microbiology. And I don't quite understand it, if you could please help thanks
A. How did you prepare the template DNA? B. Name the gene that we will be sequencing in order to identify the unknown. C. Explain why this region is considered to be the target for identification instead of sequencing the entire genome? D. What is the size (in bp) of target DNA in E. coli?
B. What do we mean by ‘conserved regions in rDNA’? Name the technique used to amplify desired target DNA and state the purpose of this amplification step. Where, in the target DNA, do primers bind during PCR (conserved vs. variable)? How many variable regions are present in rDNA? (Copy and paste the schematic diagram to show the regions)
C. Why did you run agarose gel electrophoresis? Did PCR work for both organism A and B? (Insert image of agarose gel electrophoresis here). What is the expected size of your PCR product? If you see a band in NTC, what does it imply about the results for A & B?
D. List the reagents used in preparing master mix for PCR and write one sentence about why each one was necessary.
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This is for an a 3 unknown I am doing in Microbiology.
And I don't quite...
Identification of unknown Bacteria by sequencing rDNA Having a little trouble understanding the PCR process. This...
Identification of unknown Bacteria by sequencing rDNA Having a little trouble understanding the PCR process. This is a lab we did and some homework questions regarding the sequencing rDNA to find unknown bacteria. I hope by answering these I can have better understanding of the process. The more descriptive the better. Thank you! 1. A. Name the gene that we will be sequencing in order to identify the unknown. Explain why this region is considered to be the target for...
You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR...
You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR primers that should amplify a 1 kilobase pair (kbp) PCR product that contains the gene of interest. After amplification, you will see if the PCR was successful by loading the entire reaction onto an agarose gel and performing electrophoresis to see if a product of the expected size was generated. To visualize the DNA, you will stain the gel with a fluorescent dye called ethidium bromide, which...
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Question 1. 1 2 3 4 5 6 7 8 Lane Name/Code Ladder 1B | 2B...
Question 1. 1 2 3 4 5 6 7 8 Lane Name/Code Ladder 1B | 2B 2 DNA Sample/Treatment DNA ladder Digest Digest Digest Digest Digest Digest 3B 4B 5B Negative Figure 1: 1% Super buffer agarose gel electrophoresis of restriction digest of the plasmid containing the gdhA gene insert. Based on the information above answer the following questions? 1. What ladder size used? 2. What are the two top bands and bottom bands representing? 3. Explain why the observed...
Please, I need help with this question. Show work to solve the whole question 1. You...
Please, I need help with this question. Show work to solve the whole question 1. You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR primers that should amplify a 1 kilobase pair (kbp) PCR product that contains the gene of interest. After amplification, you will see if the PCR was successful by loading the entire reaction onto an agarose gel and performing electrophoresis to see if a product of the expected size was generated. To visualize...
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Answer the following two questions with regards to the sequence
alignment of 3 individuals. Variable positions are indicated with
an *.
*1 *2 *3 AAGGGCAGCAGCAGCAGCAGCAGACTGTACCTCTTGTAAC-------AG AAGGGCAG--- ---CAGCAGTCTGTACCTCTTGTAAC-------AG AAGGGCAG- -CAGCAGTCTGTACCTCTTGTAACTCGAG Question 5 2 pts What type of variation is indicated at position *2? Question 6 2 pts TRUE or FALSE: One easy way to detect variation at position *1 among the different individual's would be to PCR amplify the position with primers to conserved flanking regions followed by gel electrophoresis. True...
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2....
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2. Suppose you begin a PCR reaction with 1 piece of double stranded DNA. After 30 cycles of replication, how many pieces of double stranded DNA do you now have? 3. What would be the consequence of having too much DNA in the sample? Would it interfere with the PCR reaction? Why? 4. What happens during the annealing step of the PCR reaction? During the...
1. If you were trying to come up with an easy-to-remember analogy for the polymerase chain...
1. If you were trying to come up with an easy-to-remember analogy for the polymerase chain reaction, you would likely say PCR is similar to a colored pencils scissors dishwasher photocopier 2. What determines the piece of DNA that is amplified in PCR? The specific primers used The source of the DNA The temperature of each cycle The specific restriction enzyme used 3. What is the role of temperature in PCR? It is used to separate and anneal the nucleic...
amplify a 161 bp fragment of the vaca gene which is only found in H. pylori....
amplify a 161 bp fragment of the vaca gene which is only found in H. pylori. You culture bacteria from gastric biopsies, extract the DNA, run the PCR, and run the gel elctrophoresis according to standard techniques. An image of the gel electrophoresis is below. 161 bp 500 bp 400 bp 300 bp 200 bp 100 bp The lanes on your gel contain the following: . M) Mass Ladder to estimate size 1) Positive Control using DNA from a lab...
Question 1. 1 2 3 4 5 6 7 8 Lane Name/Code Ladder 1B | 2B 2 DNA Sample/Treatment DNA ladder Digest Digest Digest Digest Digest Digest 3B 4B 5B Negative Figure 1: 1% Super buffer agarose gel electrophoresis of restriction digest of the plasmid containing the gdhA gene insert. Based on the information above answer the following questions? 1. What ladder size used? 2. What are the two top bands and bottom bands representing? 3. Explain why the observed...
Answer the following two questions with regards to the sequence
alignment of 3 individuals. Variable positions are indicated with
an *.
*1 *2 *3 AAGGGCAGCAGCAGCAGCAGCAGACTGTACCTCTTGTAAC-------AG AAGGGCAG--- ---CAGCAGTCTGTACCTCTTGTAAC-------AG AAGGGCAG- -CAGCAGTCTGTACCTCTTGTAACTCGAG Question 5 2 pts What type of variation is indicated at position *2? Question 6 2 pts TRUE or FALSE: One easy way to detect variation at position *1 among the different individual's would be to PCR amplify the position with primers to conserved flanking regions followed by gel electrophoresis. True...
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2. Suppose you begin a PCR reaction with 1 piece of double stranded DNA. After 30 cycles of replication, how many pieces of double stranded DNA do you now have? 3. What would be the consequence of having too much DNA in the sample? Would it interfere with the PCR reaction? Why? 4. What happens during the annealing step of the PCR reaction? During the...
1. If you were trying to come up with an easy-to-remember analogy for the polymerase chain reaction, you would likely say PCR is similar to a colored pencils scissors dishwasher photocopier 2. What determines the piece of DNA that is amplified in PCR? The specific primers used The source of the DNA The temperature of each cycle The specific restriction enzyme used 3. What is the role of temperature in PCR? It is used to separate and anneal the nucleic...
amplify a 161 bp fragment of the vaca gene which is only found in H. pylori. You culture bacteria from gastric biopsies, extract the DNA, run the PCR, and run the gel elctrophoresis according to standard techniques. An image of the gel electrophoresis is below. 161 bp 500 bp 400 bp 300 bp 200 bp 100 bp The lanes on your gel contain the following: . M) Mass Ladder to estimate size 1) Positive Control using DNA from a lab...
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