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Practical Assignment: Molecular Cloning Start Assignment Due1 Mayby23:59 Points100 Submittinga file upload Attempts0 Allowed attempts2 Available8 Apr at 9:00 - 8 May at 23:59about 1 month Molecular...

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Practical Assignment: Molecular Cloning

Start Assignment
  • Due1 Mayby23:59
  • Points100
  • Submittinga file upload
  • Attempts0
  • Allowed attempts2
  • Available8 Apr at 9:00 - 8 May at 23:59about 1 month

Molecular Project Practical Report

Molecular cloning: key steps for the production of recombinant DNAs and organisms

Total of 100 marks

Submission deadline: May 1st, 23:59

Marking scheme:

2022_ Report marking_AM 7.pptxDownload 2022_ Report marking_AM 7.pptx

Rubric

For your guidance, the marking rubric is provided below. Please post questions regarding the assessment in the discussion board as many may have the same question. We will do our best to answer discussion board questions within 24 hours (Monday -Friday). Note, questions posted on weekends may not be answered until Mondays.

Performance ratings:80-100%: HD (Excellent), 60-79%: DI (Good), 50-59%: CR (Credit) ; 50-59%: PA (Pass)

Performance descriptors: For HD XXXXXXXXXX%) you need to achieve excellently all tasks followingthe instructions and guidelines listed above

Rubric

Molecular project practical reportMolecular project practical report
CriteriaRatingsPts
This criterion is linked to a learning outcomeIntroduction
20PtsOutstandingThe introduction demonstrates a thorough understanding of the topic: experimental design and what is expected. Excellent and recent examples of the application of molecular cloning techniques in biotechnological projects with literature references.10PtsDevelopedThe introduction demonstrates a good understanding of the topic: experimental design and what is expected. The information is most relevant and supported by some primary literature references with clear attribution. Minor omissions or inaccuracies may be present.5PtsCompetentThe introduction demonstrates an adequate understanding of the topic. The information is mostly relevant but not supported by related literature references. Omissions or inaccuracies are present0PtsNot satisfactoryThe introduction is very brief and does not demonstrate an understanding of the topic.
20pts
This criterion is linked to a learning outcomeAim
5PtsOutstandingAim demonstrates an advanced and thorough understanding of the topic.2.5PtsDevelopedThe aim demonstrates an understanding of the topic.0PtsNot satisfactoryThe aim is unclear or not relevant
5pts
This criterion is linked to a learning outcomeMaterials and Methods
25PtsOutstandingMaterial and methods demonstrates a thorough understanding of the topic: Extensive description of key components of molecular cloning used in the project: enzymes, DNAs and techniques15PtsDevelopedThe interpretation shows a good understanding of the key components of molecular cloning. Contains minor omissions or inaccuracies.6PtsCompetentNot complete description of used enzymes, DNA and techniques0PtsNot satisfactoryDoes not demonstrate an understanding of using the roles of enzymes, DNA and techniques.
25pts
This criterion is linked to a learning outcomeResults
15PtsOutstandingAll results were well shown and described. Figures include a descriptive legend demonstrating high-level attention to scientific conventions.10PtsDevelopedResults were shown and described. Figure legends may be absent demonstrating limited attention to scientific conventions.5PtsCompetentResults were not well shown and described. Figures and their legends may be absent demonstrating limited attention to scientific conventions.0PtsNot satisfactoryNot all results were shown and described
15pts
This criterion is linked to a learning outcomeDiscussions
30PtsOutstandingThe interpretation of the experimental data shows a very high level and significant capability to analyse and interpret experimental results.20PtsDevelopedThe interpretation of the experimental data shows a good ability to analyse and interpret experimental results. Contains minor omissions or inaccuracies but connect to the data.10PtsCompetentThe interpretation of the experimental data shows a reasonable ability to analyse and interpret experimental results. Results and conclusions contain major omissions or inaccuracies.0PtsNot satisfactoryThe discussion is very brief and contains major omissions or inaccuracies.
30pts
This criterion is linked to a learning outcomeConclusion
5PtsOutstandingThe interpretation of the experimental data shows a very high level and significant capability to analyse and interpret experimental results.There is convincing evidence of an ability to think critically2.5PtsDevelopedShows a good ability to analyse and interpret experimental results. Contains minor omissions or inaccuracies but connects to the data.0PtsNot satisfactoryRepeating the Discussion. Not relevant and contains extensive omissions or inaccuracies.
5pts
Total points:100

Answered 1 days After May 03, 2022

Solution

Preeti answered on May 04 2022
101 Votes
Molecular Cloning
Introduction:
Molecular cloning is a technique that involves the creation of a recombinant DNA molecule. The first cloning has been performed in 1970s when the “molecular scissors” i.e., restriction enzymes were discovered (Cohen et al., 1973). Restriction enzymes are known as molecular used to cut the DNA at specific sites. A plasmid or vector is used for the transfer of foreign DNA into the other cell. The foreign gene is ligated into the genome of plasmid using restriction enzymes and DNA ligase. Due to their self-replicating nature, plasmids are used as a biological vehicle. In this study, a GFP gene, was isolated from a vector pGFP with the help of restriction enzymes and then ligated to a new vector pBC KS. After ligation, the transformation of the vector would be performed in E.coli DH5-alpha strain. Transformation is a process in which the foreign DNA is transfe
ed to bacterial cells. The E.coli cells were made competent before proceeding for transformation. There are two types of competent cells that are generally used for the transformation process- electrocompetent cells and chemically competent cells. In this study, electrocompetent cells were used for the transformation process. The screening of colonies was performed and it is expected that the positive colonies will fluorescence under the influence of blue light. The molecular cloning has become a widely used technique and has wide applications. For example, the molecular cloning of insulin (MR and KL, 1992), and the recombinant production of insulin has been a great discovery for diabetes patients. Similarly, the molecular cloning is a basic requirement for the discovery of a drug against any therapeutic target. For example, the production of a recombinant protein using molecular cloning is a first step during initial drug screening (Tian et al., 2018).
Aim
The objectives of this study are:
· Restriction digestion using HindIII and EcoRI to cut the GFP gene from pGFP vecto
· Ligation of GFP gene in expression vector, pBC KS
· Transformation of pBC KS in DH5-alpha cells
· For gene confirmation, screening of colonies was performed
Material and Methods
There are many types of molecular cloning that can be performed for the insertion of gene. In this study, the restriction enzyme cloning was performed. The stick ends were generated using restriction enzymes and the ligation of the gene to the vector was performed using DNA ligase enzyme.
Key components that are used in this study are:
1. Restriction Enzymes: HindIII and EcoRI are the two enzymes that is used in this study.
HindIII is isolated from Haemophilus influenza and...
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