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Question Description

This document indicates the format you should follow for your report.
Is important to follow the format guidelines because most journals will not accept your work if
it is too long or formatted incorrectly.

You should have a title page (if you don’t have you can lose 1 point) (which doesn’t count for
the page limit), where you have to list the names of your collaborators (lab group), highlight your
name with bold and or underline.
Catchy and informative title for your report.

Page length: Minimum 5 pages and Maximum 7 pages (if you don’t have it you can lose 1 point)
Remember, the title page and references page DO NOT count for the page length.
You may also have additional appendices at the end of your report which do not count toward
your page limit as well.

Grammar and spelling are also important for effective communication.
I could subtract up to 5 points off for bad format, bad grammar and bad spelling

Text has to be 12 -point font AND double spaced. Use a serif font (like Cambria or Times),
Margin 1-inch in both sides.

At the beginning of each section the titles should be bold and in 18-point font.

Introduction– 5 points

Should probably be around 1.5 – 2 pages in length(2 pages recomended). Basically, here you summarize IN YOUR OWN
WORDS the material in the introduction to the exercise. This is the best place to get in your
references to primary literature too. You may want to briefly talk about the history of cloning. I
highly recommend looking up terms, such as cloning methods and uses, on Wikipedia, and then
using the references in that article (you must actually go to the referenced literature and read it
though).

You should clearly describe how the experiments you are about to discuss in detail will
demonstrate the concepts you just wrote about. This does not need to be in-depth, but you should
have two or three sentences devoted to this.

The introduction is worth 5 points in total for your first report:
• 3 points for background info about the experiments (some from primary
literature). History of cloning, the various techniques
• 1 point for referencing your research adequately (your lab manual should be the
bare minimum reference here).
• 1 point for stating the purpose of this whole experiment.

make sure to provide reference

i will provide example and a template that would help (refer to experiment 3, 4,5 on the lab manual )


example(not finished not perfect)

Introduction:

DNA cloning plays an important role as a laboratory method used in molecular biology. It is simplified as the process of “isolating a DNA sequence of interest for the purposes of making multiple copies of it. This process is implemented when one cuts “a chromosome containing an interesting gene into small pieces, one of which will contain the gene and insert this gene into a plasmid shuttle, or vector, which replicates independently of the original chromosome(Lab Manual, 2020). By performing the procedure that undergoes cloning of DNA, scientists can easily isolate individual genes placed in an orgamsim to undertsnad how each gene functions and further articulate various details within them. This will ultimately help researchers have a closer understanding of our molecular biology today. Such experiments were discovered in 1968 by Arber and Linn who “succeeded in isolating an enzyme, termed a restriction factor, that selectively cut exogenous DNA”(Tirabassi, 2010). Shortly after this, Hamilton Smith expanded onto the study and confirmed Arber and Linn’s discovery by isolating a restriction enzyme from Haemophilus influenza. This major discovery plays a crucial part in the history of biology; it enhanced researchers’ ability to recombine DNA sequences and eventually grow them in different cultures. From this, we were able to familiarize ourselves with Escherichia coli, the most common bacteria used in biological laboratories.

Recombinant DNA is very much needed when the process of gene cloning is taking place. This procedure is conducted by the usage of a restriction enzyme to cut a single DNA sequence at a particular restriction site. Depending on the restriction enzyme used in the process, a cut can either be blunt or sticky. In this experiment, the restriction enzyme used was a Type II known as Hind III which formed an overhang end also known as a sticky end (Lab Manual, 2020). Once this has been conducted, the usage of a DNA ligase becomes a major part; it used to join the sticky cut end along with the vector. Now, a new recombinant DNA is created and further embedded into a host cell where the process of replication begins. If DNA fragments do not have useful restriction sites for cloning, a polymerase chain reaction (PCR) is used to help further amplify a single piece of a DNA sequence (NCBI, 2000).

FOR PARAGRAPH 3 (EDITING STILL)

The purpose of these collective experiments was to learn how to clone vectors and insert them into plasmid DNA, as well as identify if they are present by running gel electrophoresis.

In this lab, a plasmid is created through genetic engineering to be resistant to the antibiotic gentamicin. There are two types of antibiotics, one type is bacteriostatic which stops bacterial growth whereas the other type ultimately kills the bacteria. This type is known as bactericidal, one of which is gentamicin. E.Coli cultures do not contain the gentamicin resistance gene. Once the gene for resistance of gentamicin is transferred into the DNA of the E. Coli culture, it can be selected for through the use of gentamicin.

The plasmid temple is first placed in the PCR machine to produce large numbers of copies of DNA. The PCR product and isolated vector plasmid was then digested using HINDIII. Gel electrophoresis was used to separate the PCR digest, digested vector, and undigested vector based on size and charge. The insert and vector were then added to DNA ligase to form the final vector, then electroporated and incubated for the ElectroMAX DH5α-E6 cells to uptake the plasmid. To confirm transduction onto the vector plasmid, the mixture was plated onto an LB agar + gentamicin plate. Only cells with the gentamicin resistant gene would be able to grow and form colonies. Lastly, bioinformatics was used to form a genetic map of the plasmid.

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