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Now determine the sizes of the fragments generated after digesting the r-plasmid with each of the...
Now determine the sizes of fragments generated after digesting the r-plasmid with each of the restriction...
Now determine the sizes of fragments generated after digesting
the r-plasmid with each of the restriction enzymes or combinations
or restrictions enzymes.
R-plasmid digested with PstI:_______
R-plasmid digested with EcoRI:________
R-plasmid digested with PstI and EcoRI:__________
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650 4973 38mm - 45mm 55mm 59 mm...
What happens when vector DNA is digested with EcoRI and what does this tell you about...
What happens when vector DNA is digested with EcoRI and what
does this tell you about the number and distribution of EcoRI sites
in the vector?
How big (in base pairs) is (are ) the fragments generated by
digesting the vector with EcoRI?
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650...
please help me with these questions Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping...
please help me with these questions
Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping the Plasmid The first step in mapping a plasmid is to determine how many times a restriction site is found on that plasmid. Examine the results for plasmid 55 as an example. The data given in the following table are for the double digest using EcoRI and Pstl. Also, giving are the data for single digests by the individual enzymes. The numbers in the...
A genetics problem covering chapter 10 concepts. Could really use some help! The Notch gene, involved...
A genetics problem covering chapter 10 concepts. Could
really use some help!
The Notch gene, involved in Drosophila development, is contained within a restriction fragment of Drosophila genomic DNA produced by cleavage with the enzyme SalI. The restriction map of this Drosophila fragment for several enzymes (Sall, PstI, and Xhol) is shown here; numbers indicate the distances between adjacent restriction sites. This fragment is cloned by sticky-end ligation into the single Sall site of a bacterial plasmid vector that is...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp EcoRI (875) Amp(R) Psti (1550) Kan(R) Ncol (1275) 8. You will be using the plasmid shown above as a vector for a DNA cloning experiment. You will create a library of genomic DNA fragments that will then be ligated into this plasmid. The plasmid carries two antibiotic resistance genes, Amp(R) and Kan(R), an origin of replication and a LacZ gene (Lac Z alpha). The...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
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...
Cloning / Subcloning When subcloning engineering new plasmids, by inserting new DNA fragments (inserts) me plasmids...
Cloning / Subcloning When subcloning engineering new plasmids, by inserting new DNA fragments (inserts) me plasmids (now called a vector because it will carry your gene of interest) it is important to considering existing genes / DNA elements. If a site is in the middle of a gene, you could lose or destroy that gene. If there are multiple sites for an enzyme, when you paste them together, multiple possible outcomes can arise. This is undesirable, because it confounds verification...
2 Addendum 1 to this lab provides information on the location of the EcoRI, and Sspl...
2 Addendum 1 to this lab provides information on the location of the EcoRI, and Sspl cut sites in your BlueScript vector and the 1.3 kb EcoRI fragment. Using this information, (a) draw the two possible restriction maps for the recombinant pBlueKan plasmid that you constructed (see below). There are two possible maps because the 1.3 kb fragment could have been cloned in either orientation relative to the vector, Next, (b) predict the sizes of the EcoRI. and Sspl frements...
5. If you are digesting a large linear fragment that is 300,000 bp in length. How...
5. If you are digesting a large linear fragment that is 300,000 bp in length. How many fragments would be produced if the DNA was digested by the following enzymes? Assume the fragment sizes are the average sizes calculated for a random DNA sequence. Use the information in the table from Question 3. Show your work. (A) Smal (B) Haelll (C) Noti Table 4.1 Recognition Sequences and Cutting Sites of Selected Restriction Endonucleases Enzyme Alul BamHI Bgli Clal ECORI Haelll...
Now determine the sizes of fragments generated after digesting
the r-plasmid with each of the restriction enzymes or combinations
or restrictions enzymes.
R-plasmid digested with PstI:_______
R-plasmid digested with EcoRI:________
R-plasmid digested with PstI and EcoRI:__________
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650 4973 38mm - 45mm 55mm 59 mm...
What happens when vector DNA is digested with EcoRI and what
does this tell you about the number and distribution of EcoRI sites
in the vector?
How big (in base pairs) is (are ) the fragments generated by
digesting the vector with EcoRI?
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650...
please help me with these questions
Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping the Plasmid The first step in mapping a plasmid is to determine how many times a restriction site is found on that plasmid. Examine the results for plasmid 55 as an example. The data given in the following table are for the double digest using EcoRI and Pstl. Also, giving are the data for single digests by the individual enzymes. The numbers in the...
A genetics problem covering chapter 10 concepts. Could
really use some help!
The Notch gene, involved in Drosophila development, is contained within a restriction fragment of Drosophila genomic DNA produced by cleavage with the enzyme SalI. The restriction map of this Drosophila fragment for several enzymes (Sall, PstI, and Xhol) is shown here; numbers indicate the distances between adjacent restriction sites. This fragment is cloned by sticky-end ligation into the single Sall site of a bacterial plasmid vector that is...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp EcoRI (875) Amp(R) Psti (1550) Kan(R) Ncol (1275) 8. You will be using the plasmid shown above as a vector for a DNA cloning experiment. You will create a library of genomic DNA fragments that will then be ligated into this plasmid. The plasmid carries two antibiotic resistance genes, Amp(R) and Kan(R), an origin of replication and a LacZ gene (Lac Z alpha). The...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
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...
Cloning / Subcloning When subcloning engineering new plasmids, by inserting new DNA fragments (inserts) me plasmids (now called a vector because it will carry your gene of interest) it is important to considering existing genes / DNA elements. If a site is in the middle of a gene, you could lose or destroy that gene. If there are multiple sites for an enzyme, when you paste them together, multiple possible outcomes can arise. This is undesirable, because it confounds verification...
2 Addendum 1 to this lab provides information on the location of the EcoRI, and Sspl cut sites in your BlueScript vector and the 1.3 kb EcoRI fragment. Using this information, (a) draw the two possible restriction maps for the recombinant pBlueKan plasmid that you constructed (see below). There are two possible maps because the 1.3 kb fragment could have been cloned in either orientation relative to the vector, Next, (b) predict the sizes of the EcoRI. and Sspl frements...
5. If you are digesting a large linear fragment that is 300,000 bp in length. How many fragments would be produced if the DNA was digested by the following enzymes? Assume the fragment sizes are the average sizes calculated for a random DNA sequence. Use the information in the table from Question 3. Show your work. (A) Smal (B) Haelll (C) Noti Table 4.1 Recognition Sequences and Cutting Sites of Selected Restriction Endonucleases Enzyme Alul BamHI Bgli Clal ECORI Haelll...
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