Homework Answers
3)
| Gel Lane | Plasmid | Restriction Enzyme | Number of Bands Observed | Estimated Size (bp) of each band |
|---|---|---|---|---|
| 2 (tube 1) | A | N/A; uncut | 4 |
|
| 3 (tube 2) | B | N/A; uncut | 4 |
|
| 4 (tube 3) | A | BclI | 2 |
|
| 5 (tube 4) | B | BclI | 2 |
|
| 6 (tube 5) | A | EcoRI | 1 | 3.4 kb |
| 7 (tube 6) | B | EcoRI | 1 | 3.4 kb |
4)
The Uncut lane was expected to have only one band, since only one plasmid is loaded as uncut DNA. However, 4 bands are observed. This discrepancy is due to various conformations of circular DNA. Plasmids are circular DNA molecules and can, therefore, adopt various conformational states. These conformations are relaxed, coiled, supercoiled and nicked.
The Supercoiled form of Plasmid DNA migrates according to the linear size of the DNA molecule, while other coformations migrate at a slower rate, thereby showing up as separate bands on the agarose gel.
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While your gel is running, discuss your expected results for each lane on the gel •...
How did you EXPECT your uncut lane to look in the gel image? What ACTUALLY happened?...
How did you EXPECT your uncut lane to look in the gel image?
What ACTUALLY happened? What is a plausible explanation if there
was any discrepancy?
How did the observed BclI and EcoRI digest results compare to
the expected results? If they differed, list a potential reason
why?
BclI was incubated at 50°C, while EcoRI was incubated at 37°C.
Why was this necessary? What might you predict to occur if you
accidentally switched the temperatures?
3) Using the DNA ladder,...
help with questions 5 to 10 please PCB 3023L Lab #4 Protocol & Worksheet (30pt) You...
help with questions 5 to 10 please
PCB 3023L Lab #4 Protocol & Worksheet (30pt) You may work in your lab groups durine class. but all written answers must be completed individually in your own words. 1) Using the plasmid map for orientation 1 and the cDNA map as a guide, complete the plasmid map for orientation #2. (4pt) 612 1318 1 - EcoRi EcoRI Xbal ECORV -Xbal- 1662 +Bell EcoRI EcoRV Not FP -- Xhol X 2015 PRSP +...
Table 8-3. Interpretation of each lane on the gel. For Lanes 1-8, indicate the size of every DNA ...
HELP PLEASE! at least with some examples.
size of dna ladder- 1.8cm
We were unable to transcribe this imageTable 8-3. Interpretation of each lane on the gel. For Lanes 1-8, indicate the size of every DNA band on the gel by interpolating values from your standard curve. There may be multiple bands per lane.For EACH band, identify size (interpolated from the standard curve you constructed), identity, shape, and topology DISTANCE EACH BAND IN THE LANE H AS MIGRATED SIzE OF...
The PCR was a success and your target region of 770 bp in length has been...
The PCR was a success and your target region of 770 bp in length has been amplified. You now plan to digest the DNA amplicon with the restriction enzyme Eael, and clone the resulting longest fragment it into the Eael site of the 5 kb plasmid diagrammed below. 770 bp BamHI 1 200 EcoRI 800 EcoRI 4000 1000 5 kb O /1000 2000 2000 Faal You purify your recombinant plasmid from bacterial cells, and run the plasmid (uncut. or not...
Prac result Nhel Kpn I 1569 1601 Kpni 1030 2295 / 2310 Not I 988 ОТС-д...
Prac result
Nhel Kpn I 1569 1601 Kpni 1030 2295 / 2310 Not I 988 ОТС-д Sphi 2559 Nde I 484 POTC-A 6401 bp Hygro Amp Nde1 3369 pUC Ori 5260 4587 Amp: Ampicillin resistance gene 5405 - 6265 Hygro: Hygromycin resistance gene Choose the gel image with correct bands in lanes 1 to 5 to indicate the approximate positions where you expect to see these predicted DNA fragments. (1 mark) Lane 1: POTC (undigested / uncut) Lane 2: POTC-A...
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...
I need the answers for questions 2 and 3. My DNA ladder is in lane 2...
I
need the answers for questions 2 and 3. My DNA ladder is in lane 2
marked by the yellow arrow. Thanks!
Here is the only other info I have. Thanks!
Part 2: Gel purification and on Gel Slice and PCR Product Preparin modified from TBSci.com instructions for gaan A. Dissolving the Gel Stie Following electrophores, eral DNA band from grand place glice microcentrifuge tube Ib. Use an analytical balance to weigh pelice Rec die 2. Add 500 balance to...
I just need the answers to questions 2 and 3. My DNA ladder is in lane...
I just need the answers to questions 2 and 3. My DNA ladder is
in lane 2 with the yellow arrow pointing to it. Thanks!
Part 2: Gel purification and ration Gel Slice and PCR Product Preparation modified from IBSci.com instructions for gel and PCR clean-up system A. Dissolving the Gel Slice 1. Following electrophoresis, excise DNA band from gel and place gel slice in a 1.5ml microcentrifuge tube. 1b. Use an analytical balance to weigh gel slice. Record weight...
En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard...
En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard the supernatant, but keep the pellet. In step 15, you discard the pellet, but keep the supernatant. Explain why the pattern is different between the two steps and the consequence of mixing up these two steps. Procedure Part 1: mt DNA Isolation from your cheek cells. Lysis solution is used to breakdown the cells in this step, you will isolate MEONA from cheek cells....
How did you EXPECT your uncut lane to look in the gel image?
What ACTUALLY happened? What is a plausible explanation if there
was any discrepancy?
How did the observed BclI and EcoRI digest results compare to
the expected results? If they differed, list a potential reason
why?
BclI was incubated at 50°C, while EcoRI was incubated at 37°C.
Why was this necessary? What might you predict to occur if you
accidentally switched the temperatures?
3) Using the DNA ladder,...
help with questions 5 to 10 please
PCB 3023L Lab #4 Protocol & Worksheet (30pt) You may work in your lab groups durine class. but all written answers must be completed individually in your own words. 1) Using the plasmid map for orientation 1 and the cDNA map as a guide, complete the plasmid map for orientation #2. (4pt) 612 1318 1 - EcoRi EcoRI Xbal ECORV -Xbal- 1662 +Bell EcoRI EcoRV Not FP -- Xhol X 2015 PRSP +...
HELP PLEASE! at least with some examples.
size of dna ladder- 1.8cm
We were unable to transcribe this imageTable 8-3. Interpretation of each lane on the gel. For Lanes 1-8, indicate the size of every DNA band on the gel by interpolating values from your standard curve. There may be multiple bands per lane.For EACH band, identify size (interpolated from the standard curve you constructed), identity, shape, and topology DISTANCE EACH BAND IN THE LANE H AS MIGRATED SIzE OF...
The PCR was a success and your target region of 770 bp in length has been amplified. You now plan to digest the DNA amplicon with the restriction enzyme Eael, and clone the resulting longest fragment it into the Eael site of the 5 kb plasmid diagrammed below. 770 bp BamHI 1 200 EcoRI 800 EcoRI 4000 1000 5 kb O /1000 2000 2000 Faal You purify your recombinant plasmid from bacterial cells, and run the plasmid (uncut. or not...
Prac result
Nhel Kpn I 1569 1601 Kpni 1030 2295 / 2310 Not I 988 ОТС-д Sphi 2559 Nde I 484 POTC-A 6401 bp Hygro Amp Nde1 3369 pUC Ori 5260 4587 Amp: Ampicillin resistance gene 5405 - 6265 Hygro: Hygromycin resistance gene Choose the gel image with correct bands in lanes 1 to 5 to indicate the approximate positions where you expect to see these predicted DNA fragments. (1 mark) Lane 1: POTC (undigested / uncut) Lane 2: POTC-A...
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...
I
need the answers for questions 2 and 3. My DNA ladder is in lane 2
marked by the yellow arrow. Thanks!
Here is the only other info I have. Thanks!
Part 2: Gel purification and on Gel Slice and PCR Product Preparin modified from TBSci.com instructions for gaan A. Dissolving the Gel Stie Following electrophores, eral DNA band from grand place glice microcentrifuge tube Ib. Use an analytical balance to weigh pelice Rec die 2. Add 500 balance to...
I just need the answers to questions 2 and 3. My DNA ladder is
in lane 2 with the yellow arrow pointing to it. Thanks!
Part 2: Gel purification and ration Gel Slice and PCR Product Preparation modified from IBSci.com instructions for gel and PCR clean-up system A. Dissolving the Gel Slice 1. Following electrophoresis, excise DNA band from gel and place gel slice in a 1.5ml microcentrifuge tube. 1b. Use an analytical balance to weigh gel slice. Record weight...
En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard the supernatant, but keep the pellet. In step 15, you discard the pellet, but keep the supernatant. Explain why the pattern is different between the two steps and the consequence of mixing up these two steps. Procedure Part 1: mt DNA Isolation from your cheek cells. Lysis solution is used to breakdown the cells in this step, you will isolate MEONA from cheek cells....
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