Thx! 2.3.1.4.5.6.7.Part II. Your supervisors want to see the major steps you will follow to create recombinantDNA (your gene of interest inside a vector), from amplification of the gene to verifying thatyour end product doesn't have an unexpected mutation. There should be a minimum of 7steps/techniques. You must use proper and specific names for each technique.Hint: You have the DNA sample provided by the agent. What's next?Data Access Requir...Part III. You performed a PCR to amplify the open reading frame (from the start codon to thestop codon) of the Super_Toxic1 gene (plus extra 12 bps at each end containing restriction sitesto facilitate cloning). You want to run a gel to check if the PCR product contains the desiredproduct.ASne gel electrophoresis results, assuming the PCR worked perfectly. Include a DNAladder (with 50 bp, 100 bp, 150 bp and 200 bp bands), positive control, Super_Toxic1 sample,and negative control. Label each lane accordingly.B. Also indicate the expected size of the amplified fragment.

Answered: Part II. Your supervisors want to see…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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Question

Thx!

2.
3.
1.
4.
5.
6.
7.
Part II. Your supervisors want to see the major steps you will follow to create recombinant
DNA (your gene of interest inside a vector), from amplification of the gene to verifying that
your end product doesn't have an unexpected mutation. There should be a minimum of 7
steps/techniques. You must use proper and specific names for each technique.
Hint: You have the DNA sample provided by the agent. What's next?
Data Access Requir...
Part III. You performed a PCR to amplify the open reading frame (from the start codon to the
stop codon) of the Super_Toxic1 gene (plus extra 12 bps at each end containing restriction sites
to facilitate cloning). You want to run a gel to check if the PCR product contains the desired
product.
AS
ne gel electrophoresis results, assuming the PCR worked perfectly. Include a DNA
ladder (with 50 bp, 100 bp, 150 bp and 200 bp bands), positive control, Super_Toxic1 sample,
and negative control. Label each lane accordingly.
B. Also indicate the expected size of the amplified fragment.

Expert Solution

Step 1

part-3

Sure! Here are the major steps involved in creating recombinant DNA:

Isolation of the gene of interest: The first step is to isolate the specific gene of interest from the DNA sample provided by the agent. This can be achieved using techniques such as PCR (Polymerase Chain Reaction) or restriction enzyme digestion.
Amplification of the gene: Once the gene of interest has been isolated, it needs to be amplified to generate a sufficient amount of DNA for downstream applications. PCR is the most common technique used for gene amplification.
Cloning the gene: The next step is to clone the gene into a vector, which is typically a plasmid. This can be done using techniques such as ligation, where the gene of interest is inserted into the plasmid vector using DNA ligase enzyme.
Transformation: The recombinant DNA construct needs to be introduced into a host cell, such as E. coli, using a process called transformation. This involves treating the host cells with a chemical (e.g. calcium chloride) to make them competent, and then exposing them to the recombinant DNA construct.
Selection: To identify cells that have taken up the recombinant DNA, a selectable marker (e.g. antibiotic resistance gene) is included in the vector. The transformed cells are then grown in the presence of the antibiotic, which selects for cells that have the recombinant DNA construct.
Screening: Once the cells have been selected, they need to be screened to confirm that they contain the correct recombinant DNA construct. This can be done using techniques such as PCR or restriction enzyme digestion.
Verification: Finally, the recombinant DNA construct needs to be verified to ensure that it does not contain any unexpected mutations or errors. This can be done using techniques such as sequencing or restriction enzyme digestion followed by gel electrophoresis.