Transformation of E.coli with plasmid DNA & GFP purification

Transformation of E.coli with plasmid DNA & GFP purification

Write a combined report of practical class1 (transformation of E.coli) and practical class 3 (extraction of recombinant green fluorescent protein from E.coli) following the normal report format, described below.  The two classes link together and you should try to present them in a coherent integrated report.  You should decide the overall aim of the two practicals and use that as the focus for the report.

Do not include a report of practical 2 – if you do you will be penalised.

You should write the report independently and not in collaboration with a practical partner.

You are expected to follow the guidelines for the report, described below.

The length of the report should be 1500 words excluding references.

Check eVision for the submission deadline.

Guidelines for the Report
1) Title
This should be as informative as possible and relate to the aim of the work. It should be of your own composition and not simply copied from information given to you.
2)  Introduction
This should be concise and provide the reader with the relevant background of the problem.
Example 1 – in a practical report involving the denaturation of DNA, the structure of DNA should be mentioned as should denaturation and a few words about the methods used in denaturation of DNA.
Example 2 – in a practical report involving the growth of microorganisms, the use of appropriate growth media and different methods of culture would provide relevant background information.
An introduction is also the proper place to define specialist terms and abbreviations that will appear in the report eg adenosine triphosphate (ATP),  colony forming units (cfu).
The last paragraph of the introduction should state clearly the aim of the practical (but should  not be copied from the Practicals Booklet).

3)  Methods
These should be written in sentences and paragraphs, not as a list of bullet points, and in the past tense and third person.  There should be sufficient information for others to be able to repeat your work.  You should not simply copy and paste the protocols used in class.
Example 1 – “The neck of the bottle was passed through the Bunsen burner flame , and 1 cm3 of the suspension of microorganisms was  transferred to the sterile universal container” rather than  “I passed the neck of the bottle  through the Bunsen burner flame , and then I transferred 1 cm3 of the suspension of microorganisms  to the sterile universal container”.

4) Results
This section must contain a concise description of the results and a presentation of data. Numerical data are best presented in the form of tables or figures/graphs (or both as appropriate). Include images where possible – if a photograph is not possible than a hand-drawn (but neat) image is acceptable.
All images, tables, graphs etc. must be fully labelled and have a description of the results together with a brief conclusion – look at a research journal article to see what format is appropriate.

5)   Discussion
In this section do not simply describe the results – this you should have done on the Results section.
Instead you should try to analyse your results and come to some conclusions.
Practical 1
Discuss the problems involved with gene transfer by answering the questions on p12 of the Practical Booklet. Research the literature to find out how heat shock can overcome problems of gene transfer. Can heat shock be used for other methods or applications, such as those listed on p12, where gene transfer is required? How efficient is heat shock as a method of gene transfer?
Discuss the purpose of selecting bacteria by growth on different media, using your results and the questions on p18 of the Practical Booklet to guide your answer.

Practical 3
Discuss the results by answering the questions on p24 of the Practical Booklet.

6)   References
Quote any references, i.e. books, journal articles and web sites etc. that were used in writing up the report. References to support statements you may make should be cited in the text and not just in the reference list at the end of the report. Journal and text book references should be cited in the text in brackets following a particular statement and should give the author’s name and year of publication,  eg  for a journal reference “Drug resistance has become a major problem in the treatment of TB (Rieder 1994)”. The full reference giving the authors, year of publication in brackets, title of article, name of journal,  volume number and page numbers is then given in the reference list (see below).
If you use information from a text book, again you cite the author(s) and the year of publication in the text as above, and the full reference is given in the reference list (see below). eg for a text book reference,  “Hydrothermal vents support large numbers of sulphur-oxidising bacteria (Madigan et al 2000)”. The “et al” means “and others” and is used in the text when there are more than two authors, but all authors must be shown in the reference list (see below).
Book/ Journal references must be listed at the end of the report in alphabetical order according to the first author’s name (see the reference list below).
Web site references should also be cited in the text where they are given a number according to the sequence in which they appear in the text and this number is shown in brackets eg “E.coli is a Gram negative rod shaped bacillus (1) and is a common contaminant of fresh water (2)”.
Web site references are listed after journal/book references in the reference list.
Students are reminded that referenced material should be predominantly from text books and peer-reviewed research papers, rather than web sites.
Examples:
Madigan M. T., Martinko J.M. and Parker J.( 2000). Brock’s Biology of Microorganisms (9th edition). Prentice Hall International, p.354-346.
( i.e authors, year of publication , title of article, name of book, and page numbers)
Rieder, HL.(1994). Drug-resistant tuberculosis. Tuberculosis and Lung Disease. 74:324-328 (i.e. author, year of publication, title of article, name of journal,  volume number and page numbers )

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