Homework Answers
EaeI is a restriction enzyme which is named after Enterobacter aerogenes. The recognition site for this enzyme is
...YGGCCR...
...RCCGGY... Y is C or T, R is A or G
the enzyme cuts as
which result in the the production of two ends as such used for PCR
5'- GGCCR..................................................................................Y 3 '
3'-Y..................................................................................RCCGG 5'
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2. 3. Give the recognition sequence for the restriction enzyme Eael in a dsDNA sequence, indicate...
3. With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends...
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...
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...
1.When cloning a PCR product into a plasmid using restriction enzymes, the restriction enzyme recognition sequences...
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...
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...
True or false? Restriction digestion is required for PCR amplifying DNA. Ampicillin is a gene that...
True or false? Restriction digestion is required for PCR amplifying DNA. Ampicillin is a gene that encodes for ampicillin resistance. The ends produced by the endonuclease can be rejoined by a ligase, which closes the break in the hydrogen bonds formed by the complementary DNA nucleotides. Restriction recognition sites typically have a palindrome structure. During DNA ligation, DNA fragments with compatible sticky ends have a higher ligation efficiency than DNA fragments with blunt ends. PCR is a technique used to...
EcoRI is a common restriction enzyme used in cloning experiments. Its restriction sequence is G’AATTC. The...
EcoRI is a common restriction enzyme used in cloning experiments. Its restriction sequence is G’AATTC. The strand is cut at the position of the apostrophe. The 50 base pair sequence shown below contains one or more EcoRI sites. Find them and circle them. Then count the resulting size (in base pairs – bp) of the DNA fragments (pieces) left over after using EcoRI to cut this DNA sequence. Circle the EcoRI sites in the sequence below. 1- ggagaattcgctgtacgaggttaaccccgatgccATGGCATGAATTCGTG -50 List...
Suppose you digest the genomic DNA of a particular organism with the restriction enzyme SauA. Then...
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....
1. A circular plasmid has two PmeI restriction sites. A PmeI restriction enzyme will cut this...
1. A circular plasmid has two PmeI restriction sites. A PmeI restriction enzyme will cut this plasmid into two fragments. A. True B. False 2.In general, restriction enzymes that recognize four nucleotides have higher probability to produce more DNA fragments than those enzymes that recognize six nucleotides. A. true B. false 3. Which of the following sequences are palindromes? A. 5' TGGCCA 3' B. 5' GAAAAG 3' C. 5' CGATGG 3' D. 5' GACGAC 3' 4. Below are the possible...
14. A restriction enzyme (for example BamH1): (2 points) a. Is a specific sequence of DNA...
14. A restriction enzyme (for example BamH1): (2 points) a. Is a specific sequence of DNA b. Can be used to ligate DNA ends C. Recognizes palindromic DNA sequences d. Cuts random sequences of DNA
The following is a small part of a vector DNA sequence showing the NdeI and SacII...
The following is a small part of a vector DNA sequence showing
the NdeI and SacII cut sites. The
bases recognizedby the enzymes are in bold. They both are Class II
restriction enzymes. These enzymes recognize a specific sequence of
palindromic bases, i.e., reads the same on both strands
1. When you cut this DNA, shown in the figure above, by
NdeI, it will generate sticky ends. Write the two pieces
of DNA that will result after NdeI digestion and...
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...
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...
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...
True or false? Restriction digestion is required for PCR amplifying DNA. Ampicillin is a gene that encodes for ampicillin resistance. The ends produced by the endonuclease can be rejoined by a ligase, which closes the break in the hydrogen bonds formed by the complementary DNA nucleotides. Restriction recognition sites typically have a palindrome structure. During DNA ligation, DNA fragments with compatible sticky ends have a higher ligation efficiency than DNA fragments with blunt ends. PCR is a technique used to...
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....
14. A restriction enzyme (for example BamH1): (2 points) a. Is a specific sequence of DNA b. Can be used to ligate DNA ends C. Recognizes palindromic DNA sequences d. Cuts random sequences of DNA
The following is a small part of a vector DNA sequence showing
the NdeI and SacII cut sites. The
bases recognizedby the enzymes are in bold. They both are Class II
restriction enzymes. These enzymes recognize a specific sequence of
palindromic bases, i.e., reads the same on both strands
1. When you cut this DNA, shown in the figure above, by
NdeI, it will generate sticky ends. Write the two pieces
of DNA that will result after NdeI digestion and...
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