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biotechnology definition
biotechnology involves the use of living systems, organisms, or parts of organisms to manipulate natural processes in order to develop products, systems, or environments to benefit people.
what subjects does biotechnology encompass?
biology, chemistry, physics, engineering, genetics, bioinformatics, machine learning, deep learning
examples of biotechnology
manipulating DNA
recombinant DNA technology
PCR & cloning
fermentation
production of recombinant proteins
is genetic manipulation of organisms a new concept?
no; humans have been doing it for hundreds of years (i.e. breeding plants/animals in order to combine different properties)
examples of products that can created through biotechnology
recombinant proteins
recombinant plants
genetically modified animal cells
recombinant human proteins
bioethics definition
many of the products and treatments developed using biotechnology are controversial; where do you draw the moral line for your job?
steps of the scientific method
ask a question
do background research
hypothesize
experiment
collect & analyze data
report results
can hypotheses be proven?
no; an experiment can either support or reject the hypothesis
does “nothing happening” count as an observation
yes; it is still something that you noticed using your senses
the seven parts to a valid experiment are…
quantitative data
multiple replicates (≥3)
positive control
negative control
independent variables
dependent variables
constants
hypothesis
an inferred explanation of an observation or research finding: while more exploratory in nature than a theory, it is based on existing scientific knowledge.
theory
a well-substantiated and comprehensive set of ideas that explains a phenomenon in nature. a scientific theory is based on large amounts of data and observations that have been collected over time. scientific theories can be tested and refined by additional research, and they allow scientists to make predictions.
law
an expression of a mathematical or descriptive relationship observed in nature.
order of cells from smallest to largest
small molecule, virus, bacterium, animal cell, plant cell
viewed with electron microscope
less than 100µm
viewed with light microscope
greater than 1µm
average size of most cells
10µm-30µm
prokaryotic cell size
1µm
hold a microscope by the _______ & _________
base; neck/arm
methylene blue
a cationic stain (positively charged blue dye) that helps the structure inside a cell become more visible
what tool is used to measure the size of a specimen under a microscope?
micrometer
field of visions are based off of the ___X field
40
100X is ____ times smaller than 40X
2.5
400X is ____ times smaller than 40X
10
what are the three different tools used to measure volume?
graduated cylinder, serological pipet, micropipet
a graduated cylinder should be used for volumes…
greater than 10 milliliters
graduated cylinders should be read from the bottom of the….
meniscus
common graduated cylinder sizes in the lab
10mL, 25mL, 100mL, 250mL, 500mL, 1L
a serological pipet should be used for volumes….
smaller than or equal to 10mL
common serological pipet sizes in the lab
1 mL, 2mL, 5mL, 10mL
these serological pipets use blue pumps
1mL and 2mL
these serological pipets use green pumps
5mL and 10mL
serological pipets are graduated…
top to bottom & bottom to top
what does TD stand for on a serological pipet?
to delivery
volumes measured by a P-100 micropipet
10µL ≤ volume ≤ 100µL
volumes measured by a P-200 micropipet
20µL ≤ volume ≤ 200µL
volumes measured by a P-1000 micropipet
200µL ≤ volume ≤ 1000µL
solution
a mixture of two or more substances where one (solute) completely dissolves in the other (solvent)
aqueous
describing a solution in which the solvent is water
solute
the substance in a solution that is being dissolved
balance
an instrument that measures mass
weight
the force exerted on something by gravity; at sea level, it is considered equal to the mass of an object
gram
abbreviated “g”; the standard unit of mass, approximately equal to the mass of a small paper clip
solvent
the substance that dissolves the solute
molarity
a measure of concentration that represents the number of moles of a solute in a liter of solution (or some fraction of that unit)
normality
a measurement of concentration generally used for acids and bases that is expressed in gram equivalent weights of solute per liter of solution; represents the amount of ionization of an acid or base
standard balances measure…
small amounts over 900mg
analytical balances measure…
small amounts under 900mg
what is the difference between the tare button and the zero button?
in our lab, there is no difference (tare = removing the weight of a container, zero = removing any weight of any debris or air draft)
should you ever return excess of the thing you are massing back to the bottle?
no
% error =
[ (observed mass - expected mass)/expected mass ] x 100
units of measuring water
1 cubic cm = 1mL = 1g
mass/volume solutions
mass of solute/volume of solvent = final concentration (often in g/mL)
% mass/volume solutions
% value = decimal value of the g/mL
molar solutions
# of grams of solute = molarity (mol/L) x molecular weight (g/mol) x volume (L)
dilution of a concentrated solution
C1V1 = C2V2
serial dilutions
should be drawn out, each tube gets +9mL dH2O + 1 mL of the previous tube
variable
something that takes on different values
quantitative variables
can be measured numerically
qualitative variables
similar to a category; cant be measured
discrete variables
could only be a whole number (ex. 3 kids, cant be 2.89 kids)
continuous variables
could be any number (ex 1.64 seconds could be 1.64183829173081 seconds)
horizontal axis
x-axis or the abscissa
vertical axis
y-axis or the ordinate
measures of center
mean, median, mode, midrange
mean
found by adding all values and dividing by total # of values
median
the middle value when the original data is arranged in order from low to high (less affected by outliers than mean is)
mode
the value that occurs the most frequently (bimodal = 2 values) (multimodal = more than 2)
midrange
(high value + low value)/2
measures of variation
range, standard deviation
range
highest # - lowest #
standard deviation
s = sqrt [(Σ (x - x̄)²)/(n - 1)]
t-tests
can be used to determine if there is statistically significant difference between 2 data sets
null hypothesis
there is no significant difference between the two groups of data
when your p-value is less than 0.05…
reject the null hypothesis; the two data sets are significantly different
when your p-value is greater than 0.05…
accept the null hypothesis; the two data sets are not significantly different
tails
1 tail = “greater than” or “less than” (only looking for change in one direction)
2 tail = “greater than” and “less than” (looking for change in both directions)
types
type 1 = paired (before & after)
type 3 = independent, typical experiment
biological literature
any printed or electronic document written with the intent of communicating biological information
primary literature
original research results, has been never previously published, reporting their own research, new findings, always has a “materials & methods” and “data sections” (may not be labeled)
ex. journal articles, conference proceedings, dissertations/theses, patents, symposia publications, research posters
peer review
before research results are published in a scientific journal, they must pass a rigorous review process by other scientists called peer review
secondary literature
comprehensive view of previously published literature, references to primary literature, suggest new conclusions, lacks a materials and methods section, long bibliography
ex. review articles, textbooks, books/articles that interpret or review research works, any journal with “review” in the title
tertiary literature
broad overview of a topic, no materials and methods section
ex. fact books, guides and handbooks, digests, many websites
10 parts of a primary research article
title
by-line
abstract
introduction
materials & methods
results
discussion
acknowledgements
references
date of receipt/publication
UV-Vis spectrophotometer
bombards molecules with light at a particular wavelength and measures the absorbance of a cell
beer-lambert law
A = εbC
A = absorbance
ε = molar absorptivity (L/(mol cm))
b = path length (cm)
C = concentration (mol/L)
max absorption of proteins
280nm
max absorption of DNA
260nm
pure RNA ratio
~2.0
pure DNA ratio
~1.8
pure protein ratio
~0.6
purpose of DNA digestion/extraction buffer (DNA extraction)
breaks open cells and releases DNA into solution
purpose of proteinase K (DNA extraction)
digests any cellular proteins (histones), including enzymes that may break down DNA
purpose of potassium acetate (salt) (DNA extraction)
causes DNA to precipitate out of solution; allows for DNA to stay tightly bound
purpose of ice baths (DNA extraction)
further drive the precipitation of DNA from solution
purpose of centrifugation (DNA extraction)
compacts the DNA into a pellet at the bottom of the tube
purpose of isopropanol (DNA extraction)
washes the potassium acetate salts from the DNA and helps remove any remaining proteins from the solution
purpose of ethanol (DNA extraction)
removes the remaining salts, allowing DNA to return to solution in TE buffer
purpose of TE buffer (DNA extraction)
added to rehydrate the DNA for use in cuvettes
purpose of tris in TE buffer
maintains pH