Sub-cloning is a powerful technique that involves the preparation of a recombinant plasmid containing a DNA...
Sub-cloning is a powerful technique that involves the preparation of a recombinant plasmid containing a DNA fragment of interest and subsequent transformation that plasmid. In this experiment, you preformed a transformation, but the recombinant plasmid containing phage DNA was already prepared. Describe how you would make a recombinant plasmid containing a fragment you have already generated by PCR , including what enzymes you would use and the approximate length of time each step would take.
Homework Answers
- For this, we need to take plasmid DNA and the fragment of interest generated by PCR.
- Cut the plasmid DNA and fragment with same restriction endonuclease.
- Mix the two DNAs together.
- The resultant DNA will contain recombinant DNA, non-recombinant template prepared by PCR, and non-recombinant plasmid DNA.
- Through appropriate selection techniques (i.e. selectable and scorable marker), the recombinant DNA is selected from the overall mixture.
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Sub-cloning is a powerful technique that involves the
preparation of a recombinant plasmid containing a DNA...
A plasmid used as a cloning vector in E. coli must have… Does sequence similarity between...
A plasmid used as a cloning vector in E. coli must have… Does sequence similarity between genes play an important role in assigning gene function? Successful insertion of a DNA fragment into the multi-cloning region (restriction sites) of a recombinant plasmid is detected by what changes? Understand the concept of (restriction enzyme produced) DNA fragment separation by gel electrophoresis. In addition to restriction enzymes, which enzyme(s) are required to insert a fragment of DNA into a cloning vector? What is...
Which statement about gene cloning is false? A. Bacterial cells take up recombinant plasmids by transformation....
Which statement about gene cloning is false? A. Bacterial cells take up recombinant plasmids by transformation. B. Recombinant DNA molecules usually contain a DNA fragment inserted into a bacterial vector. C. To insert a gene of interest into a vector, the gene of interest and the vector must be digested with different restriction enzymes D. Transformed bacteria are plated onto media containing the appropriate antibiotic and only bacteria containing the plasmid with antibiotic resistance will grow.
1. Molecular cloning has made it possible to take a DNA fragment from nuclear DNA, containing...
1. Molecular cloning has made it possible to take a DNA fragment from nuclear DNA, containing an entire gene, and insert it into a cloning plasmid just behind a phage promoter. This means that the cloned gene can be transcribed in a test tube using a commercially available phage RNA polymerase protein. If you were to use agarose gel electrophoresis to separate RNA according to size, would the RNA produced in the test tube show any differences with RNA from...
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S...
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S pts.). The gene fragment of interest was obtained by digestion of chromosomal DNA with the restriction enzyme Sall and subsequent purification using agarose gel electrophoresis. Which antiblotic would you use in the final step of the cloning procedure, and Pst why? EcoR Sal Ampicillin Tetracycline resistanica(Ter Amp) PBR322 4,361 bp) Origin of replicatiorn (ori Pvull 8. Assume that your gene fragment from question...
please i need help with a, b, c this is the sequence 5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA...
please i need help with a, b, c
this is the sequence
5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA
AAGATCCTAACATTTTTGCGAG
TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC
AAATGGAAGTTTATATTTAAATA
GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA
ATAGTTGTGTAGATATAGGTCAT
GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA
ACAGTAATAATAACAATTTAAAACC
AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA
AACCAGAGTACAATAATAACAATT-3’
1. Polymerase chain reaction 5'- ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA ATAGTTGTGTAGATATAGGTCAT GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA ACAGTAATAATAACAATTTAAAACC AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA AACCAGAGTACAATAATAACAATT-3' a) One strand of a chromosomal DNA sequence is shown above. How would you amplify and isolate a DNA fragment defined by the sequence shown in red, using polymerase chain reaction. Design PCR primers (Forward and Reverse primers, each 20 nucleotides long, that...
1. An enzyme used to covalently join DNA segments to form recombinant DNA molecules is called...
1. An enzyme used to covalently join DNA segments to form recombinant DNA molecules is called a A. Restriction endonuclease B. Reverse transcriptase C. DNA polymerase D. Helicase E. Ligase 7. The procedure for introducing changes into specific genes is called A. An enhancer trap B. Imprinting C. RT-PCR D. DNA looping E. Gene targeting 2. Plasmids used for in vitro cloning of foreign DNA fragments are called A. Donors B. DNA chips C. Clones D. Vectors E. Conjugants 8....
8. DNA cloning. The first DNA clone using recombinant DNA technology was human insulin, which was...
8. DNA cloning. The first DNA clone using recombinant DNA technology was human insulin, which was made in 1978. The insulin gene was inserted into a plasmid and the gene was transcribed and translated from the plasmid in E coll cells. Insulin is synthesized by ribosomes in human pancreatic cells. Insulin is synthesized as a longer polypeptide, which is then trimmed by proteases to make the mature chains A and B (see Fig. 2.17 on page 39 of your textbook)....
Hi I have a problem with number 5, it involves gel analysis results. There are 2 parts, a,b,c. For A Im sure you need to make a graph with distance in (cm) on the vertical axis and log10 bp on the hor...
Hi I have a problem with number 5, it involves gel
analysis results. There are 2 parts, a,b,c. For A Im sure you need
to make a graph with distance in (cm) on the vertical axis and
log10 bp on the horitzontal. I need help figuring out where to
start and what to do. Please help!
The following question will provide practice in interpreting and analyzing gel results You obtained the DNA electrophoresis gel below. Three samples of lambda phage...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent protein Incorrect Question 4 0/0.5 pts Bacteria that did not receive a plasmid are put on an LB plate that DOES contain ampicillin. What do you expect to happen? the bacteria will create a lawn the bacteria will not grow a few colonies will be seen http:/misac.instructure.com couro/87588/quizzes/163615 7/29/2020 Review Que: BIOL-8-05-12561.202010 Transformation efficiency. Concentration of plasmid DNA. Question 8 0.5/0.5 pts Super coiled...
Chapter 10 Review: Biotechnology Learning Objectives and Application Questions Learning objectives are identified for you to...
Chapter 10 Review: Biotechnology Learning Objectives and Application Questions Learning objectives are identified for you to review your understanding of this topic. You are advised to reflect carefully on these objectives and check your own understanding to see if you have understood these essential concepts from this week’s reading. ANSWER THOROUGHLY all the application questions associated with each objective IN YOUR OWN WORDS. Learning Objective I: Evaluate the importance of biotechnology in modern societies (p. 228, 229) Application question #1...
7. Explain the procedure for cloning DNA fragment into the plasmid PBR322 (shown on the right) (S pts.). The gene fragment of interest was obtained by digestion of chromosomal DNA with the restriction enzyme Sall and subsequent purification using agarose gel electrophoresis. Which antiblotic would you use in the final step of the cloning procedure, and Pst why? EcoR Sal Ampicillin Tetracycline resistanica(Ter Amp) PBR322 4,361 bp) Origin of replicatiorn (ori Pvull 8. Assume that your gene fragment from question...
please i need help with a, b, c
this is the sequence
5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA
AAGATCCTAACATTTTTGCGAG
TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC
AAATGGAAGTTTATATTTAAATA
GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA
ATAGTTGTGTAGATATAGGTCAT
GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA
ACAGTAATAATAACAATTTAAAACC
AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA
AACCAGAGTACAATAATAACAATT-3’
1. Polymerase chain reaction 5'- ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA ATAGTTGTGTAGATATAGGTCAT GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA ACAGTAATAATAACAATTTAAAACC AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA AACCAGAGTACAATAATAACAATT-3' a) One strand of a chromosomal DNA sequence is shown above. How would you amplify and isolate a DNA fragment defined by the sequence shown in red, using polymerase chain reaction. Design PCR primers (Forward and Reverse primers, each 20 nucleotides long, that...
1. An enzyme used to covalently join DNA segments to form recombinant DNA molecules is called a A. Restriction endonuclease B. Reverse transcriptase C. DNA polymerase D. Helicase E. Ligase 7. The procedure for introducing changes into specific genes is called A. An enhancer trap B. Imprinting C. RT-PCR D. DNA looping E. Gene targeting 2. Plasmids used for in vitro cloning of foreign DNA fragments are called A. Donors B. DNA chips C. Clones D. Vectors E. Conjugants 8....
8. DNA cloning. The first DNA clone using recombinant DNA technology was human insulin, which was made in 1978. The insulin gene was inserted into a plasmid and the gene was transcribed and translated from the plasmid in E coll cells. Insulin is synthesized by ribosomes in human pancreatic cells. Insulin is synthesized as a longer polypeptide, which is then trimmed by proteases to make the mature chains A and B (see Fig. 2.17 on page 39 of your textbook)....
Hi I have a problem with number 5, it involves gel
analysis results. There are 2 parts, a,b,c. For A Im sure you need
to make a graph with distance in (cm) on the vertical axis and
log10 bp on the horitzontal. I need help figuring out where to
start and what to do. Please help!
The following question will provide practice in interpreting and analyzing gel results You obtained the DNA electrophoresis gel below. Three samples of lambda phage...
The exogenous DNA used in bacterial transformation can be, RONA mRNA molecule engineered plasmid red fluorescent protein Incorrect Question 4 0/0.5 pts Bacteria that did not receive a plasmid are put on an LB plate that DOES contain ampicillin. What do you expect to happen? the bacteria will create a lawn the bacteria will not grow a few colonies will be seen http:/misac.instructure.com couro/87588/quizzes/163615 7/29/2020 Review Que: BIOL-8-05-12561.202010 Transformation efficiency. Concentration of plasmid DNA. Question 8 0.5/0.5 pts Super coiled...
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