How to Write a Lab Report

The lab report may be a cornerstone of scientific writing, but like any other piece of writing has a Purpose,
Audience, Conventions and Trouble spots (PACT).
More information about using the PACT Principles about any section can be found in the “Living
Cell” lab manual, Appendix 1 pg 1.

The title should be concise and descriptive, states the main finding of the report in the past or present tense
- Includes full name of experiment in quotations in middle of page
- Includes full name, class, lab report number, date, and TA near bottom of page as shown
(See Appendix 1, pg 3 in “The Living Cell” lab manual)

Example:
“Micrococcus yunnanensis strain identified from WVU Life Sciences Building elevator button samples”
Rebekah Shephard
Biology 219, Lab D Report
11/3/2015
TA: Dana Huebert-Lima
Abstract:
The abstract should summarize the highlights from each section in one concise paragraph. Abstracts should
be written for a wide audience and should be no more than 250 words.
(See Appendix 1, pg 4 in “The Living Cell” lab manual

The lingering traces of viral organisms and agents in enclosed spaces can pose a threat to public health. In
order to identify potentially harmful microbes present in the WVU Life Sciences Building elevator, microbes were
isolated via swabbing on to an agar plate. After being allowed to incubate and grow, a selected colony sample was
identified phenotypically through use of gram-staining and microscopic analysis. DNA underwent centrifugation
before PCR, and its products were run on a 1% agarose gel. PCR products were then purified, quantified, and
sequenced. The sequence was obtained from a chromatogram provided by the FINCHTV program, and then
bioinfomatically analyzed on BLAST to identify the isolate as Micrococcus yunnanensis strain TGT-R9 16S ribosomal
RNA gene, partial sequence.
Introduction

The introduction should give enough background information to put the experiment in context. It should
be very broad for the general public, with each paragraph growing more detailed. You must clearly state your
hypothesis and cite all sources. *Do NOT write in the first-person.
(See Appendix 1, pg 4 in “The Living Cell” lab manual)

Microbes surround and encompass everything human beings touch and interact with at any given point, but
are too small to be seen with the naked eye. The term “microbe” is very general, and includes life forms such as
microscopic animals, protists, fungi, archaea, bacteria, and viruses (Genetic Learning Center, 2015). The problem of
concern occurs when potentially dangerous and harmful microbes, especially in the cases of bacteria and viruses, are
found in enclosed public spaces. The spread of antibiotic-resistant viruses and infectious disease has gained status in
the arena of public concern, especially during the traditional “flu season” that just happens to correlate with the
seasonal-holiday travel season. Evidence has found that microbial contamination can be removed using soap and
hot water, but must be applied with a rinsing process that would be quite difficult to enforce in small, confined
spaces such as elevators. Other studies show that while cleansing with a cloth or mop removes a majority of present
bacteria and viruses, remaining cells are also simultaneously being spread around the area by the cloth and to other
areas. (Bloomfield, SF 2007). These remaining microbes could potentially pose a threat to public health; especially
after three Ontario physicians found hospital buttons to source more unrecognized bacterial colonies than toilet
flushers. Due to the increase in antibiotic resistant microbes, unknown microbes found in enclosed spaces, such as
elevators, should be isolated and identified for researchers to accurately asses the dangers of the microbes people
unwittingly come in contact with daily.
In order to isolate such microbes from the environment, the area should be swabbed with a cotton swab
wet with pure water. The sample swabbed should be then allowed to grow on a nutrient-rich plate and
phenotypically observed. A select sample should then be microscopically analyzed and undergo gram-staining,
before it undergoes PCR to be quantified, sequenced, and identified. The 16S rRNA should be used as a “genetic
marker” in microbial identification, as it is present in almost all bacteria, its function as a necessary componenet for
protein translation, and is large enough at 1,500 bp for informatics purposes. (Abbott, et al., 2007). Furthermore,
because the 16S rRNA is a necessary ribosomal component, the rRNA is highly conserved and flank regions that
are highly variable between different bacterial families.
In this experiment, we isolated, quantified, purified, sequencesd, and identified a microbial sample isolated
from an elevator button in the West Virginia University Life Sciences Building. The microbial isolate was found to
be Micrococcus yunnanensis, a microbe taken from the roots of Polyspora axillaris, a plant native to Yunnan Province in
south-west China. (Zhao, et al., 2009).

Methods and Materials

The methodology of the lab report should chronologically describe the techniques used to complete the
experiment. Because the audience is often professional scientists, scientific terminology should be used.
Concentrations, not volumes or masses, should be noted. *Do NOT write in the first-person.

(See Appendix 1, pg 5-6 in “The Living Cell” lab manual)

Microbes were isolated from the WVU Life Sciences Building elevator by use of cotton swab and distilled
water and grown on an agar plate (Luria Broth). Plates were streaked with sterilized inoculating loops and incubated
at 37 degrees Celsius. Plates were refrigerated after one day of growth in the incubator. Selected samples of the
refrigerated colonies underwent gram staining and phenotypes were microscopically observed via oil immersion at
10X and 20X.
Cells were grown from a single colony over night at 37 degrees C. Cells were then isolated by centrifugation
at 13,000 rpm for one minute. The cell pellet formed was re-suspended in cold Solution 1 (0.5 M NaOH) and
diluted in Solution 2 (67% SDS and 0.13 M NaOH.) Solutions were incubated at room temperature for five minutes
and then diluted in cold Solution 3 (1 M Potassium Acetate, 1.67 M Acetate) before incubation on ice for ten
minutes. Samples were centrifuged for five minutes at 13,000 rpm and the supernatant was retained. The sample
containing DNA was diluted in isopropanol and incubated at room temperature before centrifuging for five
minutes at 13,000 rpm. The pellet was washed in 70% ethanol, spun for one minute at 13,000 rpm, and dried on the
bench top before being re-suspended in TE (10 mM TrisCl, 1 mM EDTA.)
The hydrated sample of genomic DNA, PCR Ready-to-go Bead, and primer mix (5uM) were incubated on
ice until ready for PCR. After a major PCR product was obtained, 6X loading dye and PCR product were run on a
1% agarose gel for 60 minutes at 100V. Nuclease free water and Membrane Binding Solution diluted the sample of
PCR product. The PCR product mixture was transferred to an SV mini-column in a collection tube and centrifuged
at 14,000 rpm for one minute. Supernatant was discarded, and the column was washed through addition of
Membrane Wash Solution diluted with 95% ethanol before being centrifuged at 14,000 rpm for five minutes.
Supernatant was again discarded before the column was centrifuged for one minute at 14,000 rpm. Nuclease free
water was added to the column before the mixture was incubated at room temperature for one minute. The column
was centrifuged at 14,000 rpm for one minute.
The concentration of double-stranded DNA and insurance of purity was observed through use of a
Nanodrop that read the absorbance at 260 nm, 280 nm, and the 260/280 ratio. A 10uL sample of DNA at 25
ng/uL was prepared for sequencing. The sample then underwent Sanger sequencing at the WVU Sequencing
Center, and nucleotides obtained from sequencing were analyzed bioinformatically using the computer program
BLAST to identify the family and species of the isolate.

Results

The results should include your data and interpretations in figures for each piece of data and a narrative tying
everything together.

(See Appendix 1, pg 6-7 in “The Living Cell” lab manual)

The experimental goal was to isolate and identify a microbial colony utilizing the sequences of the 16S
rDNA gene, which as a component of ribosomes is highly conserved and serves as priming sites for PCR
amplification of the 16S rDNA. In order to achieve such means, select microbial isolates underwent a series of
analyses and quality controls to better identify the isolated microbial species and (perhaps) family. Microbes on the
agarose plate were observed after incubation and were found to be grown three colonies of different microbes.
Colony 2 was chosen for further observation, and yielded small, flat, opaque, and yellow-white circular cocci.
Microscopic observations with oil immersion and Gram Staining were utilized to observe the unique phenotypic
traits of the microbial isolates, which were found to be Gram-Positive. Because they retained the purple color from
gram-staining, cells are indicated to have a thick peptidoglycan layer to their cell walls.

Colony # Of Colony Colony Colony Colony Colony Gra Bacteria Location of

Colonies Size Shape Edge Surface Color m Shape Swab
+/-
2 3 Small Circular Flat Opaque White-yellow Positi Circular LSB Elevator
ve

Table 1. Microbes were isolated through use of sterile technology and observed for colonial and cellular
phenotypes. Observation of the selected microbial colony revealed three different species present. Microscopic
analysis found the selected microbial isolate to be small, flat, opaque, circular in shape, and white-yellow in color. A
Gram stain showed the cell to be Gram Positive, indicating the presence of a thick peptidoglycan layer.

However, phenotypes alone were not enough to identify the microbial sample, as many different species
could be white, circular, and gram positive. After PCR, gel electrophoresis was performed to assess the presence of
a PCR product, an absolute necessity for Sanger sequencing. Major PCR products were run on a 1% agarose gel to
assess the success of the PCR reaction, and it was found that that Well 1 contained one band at 529 bp, the place
where primers were set, confirming the presence of a PCR product according to the ladder.
Nanodrop quantification was used as a quality control determine the purity and quantity of the purified PCR
product, as low 260/280 ratios indicate remaining impurities in the sample. Absorbance was read at 260 nm and
280nm and the 260/280 ratio, and 1.8 and 1.5 were found to be the readings of the respective ratios. After
undergoing Sanger Sequencing at the WVU Sequencing Center, utilizing FINCHTV to confirm that each nucleotide
base was called in each case before running the sequence through BLAST to identify the novel microbial isolate,
Micrococcus yunnanensis strain. While Micrococcus yunnanensis is found from the roots of Polyspora axillaris, a plant native
to the Yunann province of south-west China, it is possible that the plant is being grown in the fifth-floor
greenhouse of LSB and left lingering microbes in the elevator. Furthermore, the common Micrococcus luteus strain
could have undergone a mutation that led to the creation of the Micrococcus yunnanensis strain.

Discussion
The discussion should broadly describe the implications of your findings and to discuss future implications
and projects based on your data. Any limitations or mistakes should be noted here.
(See Appendix 1, pg 8 in “The Living Cell” lab manual)

In this experiment, Micrococcus yunnanensis microbes were isolated from the WVU Life Sciences Building. The
microbes were able to be phenotypically identified through gram-staining and microscopic analysis, as well through
DNA isolation, quantification, and sequencing. The presence of a major PCR product and thus the microbial isolate
was confirmed by the presence of a band at the 529 bp mark in Well 1 after running the sample on a 1% agarose
gel. Purity of the DNA was found to be ideal, as the sample had a 260/280 ratio of 1.8 and a 260/230 ratio of 1.5.
These ratios confirmed that DNA was isolated, as RNA typically has a 260/280 ratio greater than 2.0.
According to the work of Chinese researchers whom were some of first to analyze the Yunnanensis strain of
the Micrococcus genus, the strain was found to be highly gram positive, and its colonies observed to be smooth,
circular, yellow, and opaque “non-endospore-forming cocci” that ranged 0.8-1 µm in diameter. (Zhao, et al. 2009).
Such findings concur with the experimental phenotypic data, allowing for it to be assumed that no major flaws
occurred during experimentation and that the microbial isolated and sequenced is indeed Micrococcus Yunnanensis.
As so many West Virginia University students and faculty members unwittingly come in contact with
Micrococcus yunnanensis on a daily basis, the microbe’s ability to create mutated strands of influenza and the common
cold should be thoroughly examined. Further tests could include testing Micrococcus yunnanensis’s ability to take up
and transform viral DNA, and test the effects of such a virus on the human body.

Citations
The literature cited portion is the list of articles cited throughout the report.
Each journal has its own conventions for citations and can vary.(See Appendix 1, pg 9 in “The Living Cell” lab
manual)

Example: Works Cited

Abbott, SL, Janda, JM. 16S rRNA Gene Sequencing for Bacterial Identification in the
Diagnostic Laboratory: Pluses, Perils, and Pitfalls. J Clin Med. 2007;45(9): 2761-2764.
Bloomfield SF. Importance of disinfection as a means of prevention in our changing world
hygiene and home. GMS Krankenhaushyg Interdiszip. 2007;2(1): Doc25.