Homework Answers
Ans- Beta-galactosidase catalyzes the cleavage of X-gal and form 5-Bromo 4-chloro indoxyl. 5-broom 4-chloro indoxyl than dimerize and oxidize to form a bright blue insoluble pigment 5,5 dibromo 4,4 dichloro indigo.
Lac Z gene functional- blue colony, nothing inserted, no clone
only the functional Lac Z gene can metabolize X-gal.
Lac Z gene disrupted due to the insertion of the gene of interest - white colony, successfully insertion, and recombination
because the insertion site is inside the beta-galactosidase gene.
So only the white colony carries your desirable human gene, and option A white is a correct answer.
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12. If you used the PBLU plasmid as a vector to carry a desirable human gene...
3. Name a procedure which can replace the heat shock process and achieve the same goal:...
3. Name a procedure which can replace the heat shock process and achieve the same goal: 4. What is the purpose of adding ampicillin to LBA? A. LBA prevents contamination of the plates. B. Ampicillin is a growth factor that stimulates growth of E. coli cells. C. Ampicillin inhibits growth of E. coli cells that do not have amp gene. D. Ampicillin inhibits growth of E. coli cells harboring PBLU plasmid. 5. When B-galactosidase is present, X-Gal turns A. white...
Please answer these four questions. 6C. A type of plasmid used frequently as a vector in...
Please answer these four questions.
6C. A type of plasmid used frequently as a vector in recombinant DNA procedures carries gene markers for resistance to tetracycline (Tet') and to ampicillin (Amp). In one experiment, such plasmids were exposed to a restriction enzyme that cuts within the Amp' gene but leaves the Tet' gene intact. Plasmids exposed to the enzyme were next mixed with genomic restriction fragments from a cloned library of mouse DNA. The aim was to produce and detect...
Cloning/Transformation Homework Please indicate what you would see on each media plate given the following conditions...
Cloning/Transformation Homework Please indicate what you would see on each media plate given the following conditions (blue colonies or white colonies) E. coli cell Nutrient agar plate, no ampicillin, no X- gal E. coli cell Nutrient agar plate with ampicillin added, no X-gal OM OM 01 01 E. coli cell with plasmid CO ОООО Nutrient agar plate with ampicillin & X- gal added E. coli cell with plasmid and correctly inserted Insulin gene in the multiple cloning site C LB...
You have isolated a DNA fragment containing a human gene involved in lung cancer and are...
You have isolated a DNA fragment containing a human gene involved in lung cancer and are working inserting it into a standard plasmid vector used in molecular cloning which has a multiple cloning site in the lacZ gene. After cutting the DNA fragment and the vector with the same restriction enzyme and ligating the cut DNA, you transform bacteria with hopefully the recombinant vector. How do you expect to identify bacteria with the recombinant vector? A. Bacteria with the recombinant...
1)You used pBLU as the vector for the unknown insert project. After transformation, you decide to...
1)You used pBLU as the vector for the unknown insert project. After transformation, you decide to perform a blue/white test, so you plate the transformants on LB + ampicillin with X-gal. Why is ampicillin required in this plate? 2)When designing PCR primers, which primer (forward vs reverse) anneal to which DNA strand (coding vs template)?
This is what we did in the experiment .First of all,1 µl of plasmid DNA was...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
This is what we did in the experiment .First of all,1 µl of plasmid DNA was...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
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....
I don't know how to distinguish between the coding/noncoding strand on the problem. I have to put...
I don't know how to distinguish between the coding/noncoding
strand on the problem. I have to put fragment into cut segment and
then determine what happened when that occurred and why the
experimenter is receiving the observations she's getting.
Please do ligation, transcription and translation.
(IV). A cloning vector is cut with the restriction endonuclease
Sma 1, whose restriction site is
C C C (cut in between here, as shown in image) G G G
and treated with alkaline phosphatase...
explain the results for your own group and compare to other groups -especially if a group...
explain the results for your own group and compare to other
groups -especially if a group has a different result than yours ,
discuss possible errors?did the transformation occur? how do you
come to this conclusion ? did you observe the alpha complementation
or not?
what do the control plates prove to you?did you see only blue
colonies or were there any white ones as well and why?
and how to heal our results? no copy-paste, please.
this
is our...
3. Name a procedure which can replace the heat shock process and achieve the same goal: 4. What is the purpose of adding ampicillin to LBA? A. LBA prevents contamination of the plates. B. Ampicillin is a growth factor that stimulates growth of E. coli cells. C. Ampicillin inhibits growth of E. coli cells that do not have amp gene. D. Ampicillin inhibits growth of E. coli cells harboring PBLU plasmid. 5. When B-galactosidase is present, X-Gal turns A. white...
Please answer these four questions.
6C. A type of plasmid used frequently as a vector in recombinant DNA procedures carries gene markers for resistance to tetracycline (Tet') and to ampicillin (Amp). In one experiment, such plasmids were exposed to a restriction enzyme that cuts within the Amp' gene but leaves the Tet' gene intact. Plasmids exposed to the enzyme were next mixed with genomic restriction fragments from a cloned library of mouse DNA. The aim was to produce and detect...
Cloning/Transformation Homework Please indicate what you would see on each media plate given the following conditions (blue colonies or white colonies) E. coli cell Nutrient agar plate, no ampicillin, no X- gal E. coli cell Nutrient agar plate with ampicillin added, no X-gal OM OM 01 01 E. coli cell with plasmid CO ОООО Nutrient agar plate with ampicillin & X- gal added E. coli cell with plasmid and correctly inserted Insulin gene in the multiple cloning site C LB...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
This is what we did in the experiment .First of all,1 µl of
plasmid DNA was added into the tube which contains competent cells
and the tube was tapped gently to mix DNA and the competent
bacteria. After that it was placed on ice for 30 minutes. Then, the
tube with the competent bacteria and plasmid were transferred to
heating block at 42 °C and the tube was leaved in there exactly 90
seconds. 0.25 ml of LB broth was...
I don't know how to distinguish between the coding/noncoding
strand on the problem. I have to put fragment into cut segment and
then determine what happened when that occurred and why the
experimenter is receiving the observations she's getting.
Please do ligation, transcription and translation.
(IV). A cloning vector is cut with the restriction endonuclease
Sma 1, whose restriction site is
C C C (cut in between here, as shown in image) G G G
and treated with alkaline phosphatase...
explain the results for your own group and compare to other
groups -especially if a group has a different result than yours ,
discuss possible errors?did the transformation occur? how do you
come to this conclusion ? did you observe the alpha complementation
or not?
what do the control plates prove to you?did you see only blue
colonies or were there any white ones as well and why?
and how to heal our results? no copy-paste, please.
this
is our...
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