If you have plasmid DNA and want to isolate a specific gene, do you need to...
If you have plasmid DNA and want to isolate a specific gene, do you need to perform both RFLP and PCR? Or would only one of those techniques be needed?
Homework Answers
Only Polymerase Chain Reaction (PCR) technique is sufficient to isolate specific gene from the plasmid DNA.
Polymerase Chain Reaction (PCR) amplifies specific region in the DNA strand by using primers (in our scenario plasmid DNA region is target where specific gene is present).
RFLP technique is used for identification of variations/polymorphisms in the homologous DNA sequences, so this technique is not used in isolation of specific gene from the plasmid DNA.
Add Answer to:
If you have plasmid DNA and want to isolate a specific gene, do
you need to...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circul...
10 please and 7
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circular DNA frequently found in prokaryotes. Aside from being smaller, how is it different from the prokaryotic genome? You place equal amounts of plasmid DNA in 4 different tubes and incubate the DNA with increasing amounts of the enzyme topoisomerase I for 1 hour (0 enzyme units 0.25 enzyme units, 0.5 enzyme units and 1 enzyme unit). You then analyze the plasmid...
I need help with both parts a and b of isolation and amplification. I would like...
I need help with both parts a and b of isolation and amplification. I would like both parts answered since it only counts as one question. Thank you. Gene is SCN5A Gene Isolation and amplification of an exon (>100 bp) of a gene by PCR (5 points) a) Isolate human genomic DNA b) Design PCR primers to amplify the exon and add restriction sites to both 5’ and 3’ of the DNA
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...
I didnt do quite as well as i wpuld have liked on my molecular bio exam....
I
didnt do quite as well as i wpuld have liked on my molecular bio
exam. Please help me reciew what I missed. Thanks!
We were unable to transcribe this imageWe were unable to transcribe this imageA fragment restricted with EcoRI enzyme can be used for ligation into a plasmid that was restricted with BamHI because both the insert and the plasmid contain sticky ends a True b) False 10. Polynucleotide probes can be used to screen a genomic library...
BTH2732 Recombinant DNA Technology For answer all questions below part, Ignore question 1. Just do question...
BTH2732 Recombinant DNA Technology
For answer all questions below part, Ignore question 1. Just
do question 2 and below
Part A: Refer to Lecture 3 & Supplementary Video: https://www.youtube.com/watch?v SiwNtQYLKeU Virtual cloning of the Human p53 gene You are required to devise a set of practical methodologies in order to carry out a relatively simple molecular biology task. 1. You must identify the steps required 2. Formulate a set of practical protocols needed to carry out those steps . Consider...
You isolate three E. coli mutants that are unable to synthesize histidine. You amplify the gene...
You isolate three E. coli mutants that are unable to synthesize histidine. You amplify the gene responsible for the phenotype from the wildtype (WT) and mutant strains (1, 2, and 3) using PCR and run the PCR products on an agarose gel. Based on the results, which mutant is likely to have an deletion in the gene of interest? (The top of the gel is where the DNA was loaded - the negatively charged end) Wt 1 2 3
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To...
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To begin the recombinant DNA process, scientists must first ide at codes for the production of the protein they want to manufacture. One is to go backwards from the amino acid sequence of the desired protein to ide sequence of the gene. After scientists have identified the gene, they m it. Restriction enzymes or endonucleases from bacterial cells are key in th ia produce restriction...
You hypothesize that a DNA sequence located 3.0 kb upstream of the promoter of a gene...
You hypothesize that a DNA sequence located 3.0 kb upstream of the promoter of a gene you are studying contains an enhancer. To test your hypothesis, you use recombinant DNA techniques to insert a DNA fragment containing this DNA sequence into a plasmid also containing a “reporter gene.” Note:A reporter gene encodes for a protein that you can detect when expressed. For example, you could use the lacZ gene that encodes for the enzyme β-Galactosidase as a reporter gene. As...
You have cloned the Pepsi Gene into the vector pKEN. You used two restriction enzymes, BamHI...
You have cloned the Pepsi Gene into the vector pKEN. You used two restriction enzymes, BamHI and EcoRI to force the cloning in a directional way. The plasmid (pKEN) is ~5 kb long and the insert Pepsi 800bp long. After transformation into E. coli you grow up several colonies and isolate the vector. After digestion of the vector it looks like you have three potential clones. You need to sequence the gene to make sure you have the correct clone....
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
10 please and 7
You isolate plasmid DNA from bacteria (Questions 7-10) 7) A plasmid is an extrachromosomal circular DNA frequently found in prokaryotes. Aside from being smaller, how is it different from the prokaryotic genome? You place equal amounts of plasmid DNA in 4 different tubes and incubate the DNA with increasing amounts of the enzyme topoisomerase I for 1 hour (0 enzyme units 0.25 enzyme units, 0.5 enzyme units and 1 enzyme unit). You then analyze the plasmid...
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...
I
didnt do quite as well as i wpuld have liked on my molecular bio
exam. Please help me reciew what I missed. Thanks!
We were unable to transcribe this imageWe were unable to transcribe this imageA fragment restricted with EcoRI enzyme can be used for ligation into a plasmid that was restricted with BamHI because both the insert and the plasmid contain sticky ends a True b) False 10. Polynucleotide probes can be used to screen a genomic library...
BTH2732 Recombinant DNA Technology
For answer all questions below part, Ignore question 1. Just
do question 2 and below
Part A: Refer to Lecture 3 & Supplementary Video: https://www.youtube.com/watch?v SiwNtQYLKeU Virtual cloning of the Human p53 gene You are required to devise a set of practical methodologies in order to carry out a relatively simple molecular biology task. 1. You must identify the steps required 2. Formulate a set of practical protocols needed to carry out those steps . Consider...
You isolate three E. coli mutants that are unable to synthesize histidine. You amplify the gene responsible for the phenotype from the wildtype (WT) and mutant strains (1, 2, and 3) using PCR and run the PCR products on an agarose gel. Based on the results, which mutant is likely to have an deletion in the gene of interest? (The top of the gel is where the DNA was loaded - the negatively charged end) Wt 1 2 3
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To begin the recombinant DNA process, scientists must first ide at codes for the production of the protein they want to manufacture. One is to go backwards from the amino acid sequence of the desired protein to ide sequence of the gene. After scientists have identified the gene, they m it. Restriction enzymes or endonucleases from bacterial cells are key in th ia produce restriction...
Most questions answered within 3 hours.
-
Dealing with conflict, in my opinion, is a skill and an art. I
believe that those...
asked 7 minutes ago -
6. A long straight wire with circular cross section and radius
1.5 cm carries a 1.2...
asked 11 minutes ago -
Deacon Corporation has provided the following financial data
from its balance sheet and income statement:
Year...
asked 1 hour ago -
List and discuss at least three variables from everyday life for
which you expect the variable ...
asked 59 minutes ago -
A high-pass filter consists of a 1.60 μF capacitor in series
with a 125 Ω resistor....
asked 1 hour ago -
A newly developed artificial intelligence has trouble
distinguishing hand-written numbers from hand-written alphabets.
There are pictures...
asked 1 hour ago -
The carbonate ion CO3 2- has multiple resonance structures.
a. Draw them and assign the hybridization...
asked 1 hour ago -
In doing an allocation for a basket purchase
Use the individual costs and the total market...
asked 1 hour ago -
what were the social political and economic causes and impacts of
the late 19th century urban...
asked 1 hour ago -
if the magnitude of a velocity vector is 10m/s and it
makes an angle of 45...
asked 1 hour ago -
Which amino acid has a basic R group?
A) tyrosine
B) Isoleucine
C) Glutamic Acid
D)...
asked 1 hour ago -
Is there the need for the government to have hackers. Please
share your thoughts.
asked 1 hour ago