Describe ligation reaction. Discuss the possible products of ligation reaction.What is the maximum transformation efficiency that...
Describe ligation reaction. Discuss the possible products of ligation reaction.What is the maximum transformation efficiency that can be expected? What things can influence the transformation efficiency and ligation reaction using 2 endonuclease for subcloning instead of one?
Homework Answers
Ligation
The desired gene of interest irrespective of species, can be expressed (as protein ) in large quantity by harnessing the E. coli bacterium as host with help of recombinant DNA technology.
A process used to join two DNA fragments in molecular cloning process by formation of phosphodiester bonds.
The product of cloning results in the clone ( gene of interest DNA+ vector DNA), which can be transformed into E. coli for expression of our gene of interest.
The transformation efficiency can be defined by number of colony forming unit (cfu) can be given by the formula
Transformation efficiency =colonies found in the plate / concentration of DNA used/ dilution of DNA used for plating.
Factors affecting transformation
- Bacterial strains used
- Growth of the bacterium
- Media composition of the medium used for bacterial growth
- concentration of the DNA used
The choice of using 2 endonucleases while sub-cloning inorder to excise the desired DNA fragment fully from the vector DNA. The ligation reaction joins the two nicks present in both gene of interest and vector DNA.
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Describe ligation reaction. Discuss the possible products of
ligation reaction.What is the maximum transformation efficiency
that...
Describe ligation reaction. Discuss the possible products of ligation reaction. What happened to e coli that did not take any the vector? Drug resistance? What is the maximum transformation efficiency...
Describe ligation reaction. Discuss the possible products of ligation reaction. What happened to e coli that did not take any the vector? Drug resistance? What is the maximum transformation efficiency that can be expected? What things can influence the transformation efficiency and ligation reaction using 2 endonuclease for subcloning instead of one?
Describe the ligation reaction. Discuss the possible products of the ligation reaction. Which ones would you...
Describe the ligation reaction. Discuss the possible products of the ligation reaction. Which ones would you expect to find in your plates? 2. Compare the ligation and control plates. Are the plates different/similar? Why? Are the transformation efficiencies similar or different? Why? What happened to e coli that did not take any the vector? Drug resistance? What is the maximum transformation efficiency that can be expected? How this compares to the trasformation efficiency of your plates? 3. For the the...
the options are with a : - Name all possible unwanted products of the ligation reaction...
the options are with a : -
Name all possible unwanted products of the ligation reaction you will be performing in the lab. The reaction will involve incubating blunt-ended plasmid and blunt- ended PCR product with DNA ligase and ATP. -primer dimers -self-ligation of inserts -ligation of insert with primers -ligation of primers with vector -ligation of vector with 2 inserts -self-ligation of vector
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There are three possible products of this reaction. Draw all of the possible oxidation products. Consider...
There are three possible products of this reaction. Draw all of
the possible oxidation products. Consider (a) primary alcohols can
be oxidized to either an aldehyde or to carboxylic acid (b) both
alcohols may be oxidized, or only one.
NaOCI OH CH,COOH
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Maximum points possible in parenthesis It has been suggested that the term ozon describe ozone in the stratosphere. List/discuss at least one a of each term? (2pts) the term ozone screen would be a better phrase than ozone layer to scuss at least one advantage and one disadvantage Ηταννα
1. Balance: Describe the reactants. Describe the products. What is the evidence of reaction? Classify by...
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a) show the reaction mechanism for this condensation reaction b) of all the possible products there...
a)
show the reaction mechanism for this condensation reaction
b) of all the possible products there is one dominant, which
one and why? And what will work as a nucleophile?
Bas
2. Draw two possible products for the reaction shown below. Are your products formed as a...
2. Draw two possible products for the reaction shown below. Are your products formed as a racemic mixture? Which of your two products do you expect to be the major one? Why? 1 equiv. Br2 THF possible product 1 possible product 2 Possible product 1 a racemic mixture? Yes No Possible product 2 a racemic mixture? Yes No Which would be major? Why?
2. Draw two possible products for the reaction shown below. Are your products formed as a...
2. Draw two possible products for the reaction shown below. Are your products formed as a racemic mixture? Which of your two products do you expect to be the major one? Why? 1 equiv. VON Br2 THE possible product 1 possible product 2 Possible product la racemic mixture? Yes No Possible product 2 a racemic mixture? Yes No Which would be major? Why?
the options are with a : -
Name all possible unwanted products of the ligation reaction you will be performing in the lab. The reaction will involve incubating blunt-ended plasmid and blunt- ended PCR product with DNA ligase and ATP. -primer dimers -self-ligation of inserts -ligation of insert with primers -ligation of primers with vector -ligation of vector with 2 inserts -self-ligation of vector
There are three possible products of this reaction. Draw all of
the possible oxidation products. Consider (a) primary alcohols can
be oxidized to either an aldehyde or to carboxylic acid (b) both
alcohols may be oxidized, or only one.
NaOCI OH CH,COOH
Maximum points possible in parenthesis It has been suggested that the term ozon describe ozone in the stratosphere. List/discuss at least one a of each term? (2pts) the term ozone screen would be a better phrase than ozone layer to scuss at least one advantage and one disadvantage Ηταννα
1. Balance: Describe the reactants. Describe the products. What is the evidence of reaction? Classify by the type of reaction. 2. Balance: CuSO4 (aq) + Zn (s) -→ Znso4 (aq) + Cu (s) Describe the reactants. Describe the products What is the evidence of reaction? Classify by the type of reaction. CuCl2 (ag)+ 3. Balance: HCl (a)+CuSO4 (ag) Describe the reactants. Describe the products What is the evidence of reaction? Classify by the type of reaction.
a)
show the reaction mechanism for this condensation reaction
b) of all the possible products there is one dominant, which
one and why? And what will work as a nucleophile?
Bas
2. Draw two possible products for the reaction shown below. Are your products formed as a racemic mixture? Which of your two products do you expect to be the major one? Why? 1 equiv. Br2 THF possible product 1 possible product 2 Possible product 1 a racemic mixture? Yes No Possible product 2 a racemic mixture? Yes No Which would be major? Why?
2. Draw two possible products for the reaction shown below. Are your products formed as a racemic mixture? Which of your two products do you expect to be the major one? Why? 1 equiv. VON Br2 THE possible product 1 possible product 2 Possible product la racemic mixture? Yes No Possible product 2 a racemic mixture? Yes No Which would be major? Why?
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