The insert is cloned into the ApaI enzyme site.
Question 4 C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2:...
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...
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...
Please help. Every time I have posted this I have gotten a different answer. Draw the plasmkd containing the cloned insert Indicate clearty where the insert will be ocated. Include RE sites and distances. 300 EcoR3400 544kb 1000 2000 Cloned Insert The PCR was a success and your target region of 440 bp in length has been ampiied You igate a short linker containing an Apal restriction enzyme site onlo both ends of the PCR product, digest it with Apal...
3. 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)? (4) GGCCR PCR amplicon RCCGG 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...
1.When cloning a PCR product into a plasmid using restriction enzymes, the restriction enzyme recognition sequences in the PCR product most likely came from _______, and the restriction enzyme recognition sequences in the plasmid most likely came from ________. a. A multiple cloning site / the primers b. The primers / a multiple cloning site c. Both came from primers d. Both came from the multiple cloning site e. Naturally present in the gene of interest / the multiple cloning...
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...
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...
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
PART 3 -SCREENING POTENTIAL CLONES (2 MARKS). After completing a cloning experiment, we screen potential clones to determine which ones are successful. The experiments outlined in Session 2-4 in the lab manual describe how we do this for the LEAPS plasmid. In this experiment using just the EcoRI site to facilitate cloning we have an additional problem - both ends of the PCR product have an EcoRI site and this means that the amplified sequence can be cloned into the...
Suppose you digest the genomic DNA of a particular organism with the restriction enzyme SauA. Then you ligate the resulting fragment into a unique BamHI cloning site of a plasmid. The sequence of the restriction sites and position of cleavage is shown below Note: X and Y and their complementary bases Z and Y’ respectively can be any base (A,C, G, or T) 1) As you can see, ligation is possible because the two restriction enzymes produce compatible sticky ends....