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BIOL 223 Experiment 1 - Lab Report Writing Guidelines Fall 2024

The document outlines the procedures and requirements for Experiment 2 at Trinity Western University, focusing on plasmid DNA characterization through restriction digests and sequence analysis. It includes instructions for presenting results in tables and figures, analyzing gel electrophoresis data, and discussing findings related to plasmid size and GFP sequence confirmation. Additionally, it emphasizes the importance of proper referencing and acknowledges contributions from others in the lab.

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11 views4 pages

BIOL 223 Experiment 1 - Lab Report Writing Guidelines Fall 2024

The document outlines the procedures and requirements for Experiment 2 at Trinity Western University, focusing on plasmid DNA characterization through restriction digests and sequence analysis. It includes instructions for presenting results in tables and figures, analyzing gel electrophoresis data, and discussing findings related to plasmid size and GFP sequence confirmation. Additionally, it emphasizes the importance of proper referencing and acknowledges contributions from others in the lab.

Uploaded by

kaivanminab
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Trinity Western University

Experiment 2

Plasmid DNA characterization using restriction


digests and sequence analysis
This lab activity was taken from the Ahern manual which has been placed on Moodle for you to use as a reference. Be
sure to properly reference any information you glean from this manual or any other source you find!

RESULTS
Table 1 should consist of the results produced in the first lab for the concentration and A260/A280 of the DNA
samples. Not all students will have average values; reporting the exact values are sufficient.

Figure 2 should consist of the gel produced in the second lab. You must follow the exact format of presenting your
gel as labelled below.
*Note: Gels should always be formatted as per the following example

Figure 2 Gel electrophoresis of pUN DNA digested with two different exonucleases, and electrophoresed in a 1.2%
agarose gel
Trinity Western University

Always remember to refer to bands specifically (ie. In Figure 2, Lane 2, there are 2 bands of approximately
____bp and ___bp in size.)

Your molecular weight standard sizes can be identified using this:

To analyze your gel – choose one of the two options: ​


Distance migrated using ImageJ: Open gel image in ImageJ and use the line tool to determine the distance from the
front of the well to the front of the band. Under ‘Analyze’, select ‘Measure’ to find out the distance in pixels. This
unit is okay to use on your lab report so long as it is indicated on your Tables and Figures.​
Distance migrated using ruler: Print off the gel image onto a plain piece of paper and measure the distance migrated in
mm using a ruler. Remember to indicate the correct unit in your Tables and Figures.

Table 2 should contain the migration data from Figure 2 for your molecular weight standards. (Table 2.3 in the
Experiment 1 Part 2 Manual)

Table 3 should contain the migration data from Figure 2 for plasmid pUN bands. (Table 2.4 in the Experiment 1 Part
2 Manual). Use the distance of the standard bands migrated to fill in the bp size of your plasmid bands.
Approximation is fine.
Trinity Western University

To determine the size of the cut and uncut plasmid bands, compare the distance that your bands migrated vs. the
distance the standard bands migrated. For example, if your 8000bp standard band migrated 5mm and your plasmid
band migrated about 4.8mm, you can approximate to say that your plasmid band is about 8000bp. Approximated
value is sufficient here.
Trinity Western University

Table 4 should contain BLAST Results.


Match Description E value Identities [X/Y (%)]

What is the size of the jellyfish GFP sequence? What is the size of your GFP sequence? Is there a
difference between the two sizes? ​

DISCUSSION (not an exhaustive list, all guidelines as stated in class still apply)

-​ pFPV25.1 is 5395bp in size. Do your restriction enzyme digest results show that your plasmid is the same size
or a different size? Explain your reasoning.

-​ Did you confirm that your plasmid contains the GFP sequence using your analysis methods? Why or why
not?

-​ For Figure #2: You need to speak about the quality of the gel produced (refer to the document on Moodle
that deals with analyzing the quality of your gel). You should also discuss the banding pattern and the
significance between the cutting of the plasmid DNA by the three different restriction enzymes. Is there
anything about the gel that was unexpected?

-​ Is this approximation method for estimating band sizes accurate? Is there a more accurate method that you
could use?

-​ Include any references you used, but the reference section will not be worth any marks (previously graded in
the last part).

ACKNOWLEDGEMENTS
Must include all people who helped you with this lab (sharing enzymes or DNA) etc.

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