Homework Answers
Restriction enzyme can create fragments with sticky ends and they have unpaired bases at 3' and 5' regions
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3. With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends...
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...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant plasmid containing the PCR fragment with the different restriction enzymes as listed. Recombinant plasmid Restriction enzyme No. of cuts Fragment sizes EcoRI BamHI Eael EcoRI + BamHI +Eae 1 -teamHI EcoRI 4000 EcoRI 6k6 1000 2000 es Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictch enzyme Egel, and clone the resulting longest fragment into...
2. 3. Give the recognition sequence for the restriction enzyme Eael in a dsDNA sequence, indicate...
2. 3. Give the recognition sequence for the restriction enzyme Eael in a dsDNA sequence, indicate the cleavage sites. (2) With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends of the fragment that will be generated after digestion with Eael (indicate these in the two blocks below). What type of ends are generated by this enzyme (blunt/sticky, 5' or 3' overhang)? PCR amplicon بیا بیا
Question 4 C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2:...
Question 4
C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2: Restriction enzyme digest analysis The PCR was a success and your target region of 440 bp in length has been amplified. You igate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below Bamll 300 EcoRI 3400 1000 2000 5...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, dige...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
Question 2,3 and 4 aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A sel...
Question 2,3 and 4
aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A selectable marker -Drug resistant gene 1. 2. Draw the end of the fragment that will be generated after digestion with the Apal enzyme. Indicate the type of ends generated by this enzyme (Blunt/Sticky, 5' or 3' overhang) (3) 3. What is the size of the plasmid...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictich enzyme Egel, and done the resulting longest fragment into Eael site of 5 KB plasmid. 4. You now run the four RE digests of the recombinant plasmid from question (4) on an agarose gel and stain it with ethidium bromide (EtBr). Indicate your expected profile on the gel below. Clearly indicate the...
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...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb)....
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
Can someone please help me answer these? Neat work is greatly appreciated. Explanation will also be...
Can someone please help me answer these? Neat work is greatly
appreciated.
Explanation will also be appreciated (optional, if you have
time).
Thank you so much! I will give a big thumbs up :)
(1) You start with the pUC18 plasmid that is a -2.7 kb circular DNA molecule. The multiple cloning site (i.e., polylynker) has unique restriction sites in the following order: HindIII, BamHI, EcoRI. Recall that unique restriction site means that these sites are only found once on...
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...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant plasmid containing the PCR fragment with the different restriction enzymes as listed. Recombinant plasmid Restriction enzyme No. of cuts Fragment sizes EcoRI BamHI Eael EcoRI + BamHI +Eae 1 -teamHI EcoRI 4000 EcoRI 6k6 1000 2000 es Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictch enzyme Egel, and clone the resulting longest fragment into...
2. 3. Give the recognition sequence for the restriction enzyme Eael in a dsDNA sequence, indicate the cleavage sites. (2) With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends of the fragment that will be generated after digestion with Eael (indicate these in the two blocks below). What type of ends are generated by this enzyme (blunt/sticky, 5' or 3' overhang)? PCR amplicon بیا بیا
Question 4
C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2: Restriction enzyme digest analysis The PCR was a success and your target region of 440 bp in length has been amplified. You igate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below Bamll 300 EcoRI 3400 1000 2000 5...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
Question 2,3 and 4
aBbCcDdE AaBbCoDdE Normal No Spacing BamHI 300 EcoRI 3400 1000 2000 5 kb EcoRU Apal List three components a cloning vector must contain A cloning site A replication origin A selectable marker -Drug resistant gene 1. 2. Draw the end of the fragment that will be generated after digestion with the Apal enzyme. Indicate the type of ends generated by this enzyme (Blunt/Sticky, 5' or 3' overhang) (3) 3. What is the size of the plasmid...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictich enzyme Egel, and done the resulting longest fragment into Eael site of 5 KB plasmid. 4. You now run the four RE digests of the recombinant plasmid from question (4) on an agarose gel and stain it with ethidium bromide (EtBr). Indicate your expected profile on the gel below. Clearly indicate the...
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...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
Can someone please help me answer these? Neat work is greatly
appreciated.
Explanation will also be appreciated (optional, if you have
time).
Thank you so much! I will give a big thumbs up :)
(1) You start with the pUC18 plasmid that is a -2.7 kb circular DNA molecule. The multiple cloning site (i.e., polylynker) has unique restriction sites in the following order: HindIII, BamHI, EcoRI. Recall that unique restriction site means that these sites are only found once on...
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