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Biotechnology on the
Cheap- Hands on Labs for
Under $50Jessie Dorman
Dilution Labs
Dye Lab- (Serial Dilution) Use Methylene Blue, tap water, microtiter plate (I use 12
well plates) Can be done using cups or tubes
Bleach Lab (C1V1=C2V2) Bleach’s caustic effects on Jeans Jeans, tap water, bleach, graduated cylinder, cups,
forceps, timer NEW LAB- What concentration do you need to be
effective? (Serial Dilution) Use Distilled water, bleach, tubes (with caps), petri-plates,
culture broth, E. Coli, Loop-needle OR dropper NOT posted on website due to copy right (U. of Missouri):
Google Microbes in Action.
Bacterial Growth
Hand wash Lab Soap, hand-sanitizer, third option (wipes?), petri-
dishes Disinfectants Lab
Disinfectants (bleach, H2O2, Alcohol, Lysol, etc.), water, petri-dishes, spoiled milk (could use E. coli), cups (or well plate), forceps
You Don’t Know Where That’s Been Supplies- Vary depending on student design
(see possible list on supplies handout or on activity handout)
Optional Bacteria Lab
Similar to You Don’t Know but instead of swabbing surfaces students could use soil, water, and air samples.
Simply add soil and water samples to start a broth culture (usually ~5-10 mL sample to ~10 mL broth), then incubate for 24-48 hours (until culture gets cloudy). Works best if the culture is “agitated” a few times during the first few hours.
Then simply swab the culture onto petri-dishes Air samples, expose broth to the air sample for
an amount of time and then do the same as you did for the soil and water samples.
ELISA
ELISA is an “Enzyme Linked ImmounoSorbant Assay
Purpose is to determine if an antibody protein is present in the serum and if so how much.
The protein being present indicates and immune response occured
The protocol obtained is from Carolina and only looks for positive (protein present), negative(protein not present), and slight positive (low amounts of protein present)
ELISA Cont.
The Carolina lab uses patented “enzymes” to do the reaction. It is a bit more specific then my Cheap Version of it.
Need Distilled Water, base, phenolphthalein, 1.5 mL tubes
My way, while less specific as mentioned above, is considerably cheaper. (Even with buying all of the big items such as the 1.5 mL tubes, and pipettes it only costs ~$98 and those items are useful in many different experiments.)
The Kit from Carolina is ~$105 with kit refills (“enzymes” costing ~$46 which need to be ordered within a few weeks of doing the experiment, frozen on receipt and replaced each year).
Carolina Protocol
ELISA Notes of Interest
If using my version be sure to “neutralize” all items (pipettes, tubes, plates) used or when you go to do the experiment again you may get “false positives” or results that show before adding the Chromogen (Phenolphthalein)
I just use a bin of water with a quarter to half cup of lemon juice
Another way to do the experiment is to do the dilution series down the plate. This is how it is done “in the real world”. If doing that you will need to use the 96 well plates and essentially do 1:2 dilutions down the plate (ex have 10 uL of diluent and add an mix 10 uL of the previous well’s sample.
See file named
“ELISA Procedure for Measuring Serum Antibody Titer”
Antibody Titer Method1 2 3 4 5 6 7 8 9 10 11 12
A S1 100
S1 100
S1 100
S2 100
S2 100
S2 100
S3100
S3100
S3100
S4100
S4100
S4100
B S1 50
S1 50
S1 50
S2 50
S2 50
S2 50
S3 50
S3 50
S3 50
S4 50
S4 50
S4 50
C S125
S125
S125
S225
S225
S225
S325
S325
S325
S425
S425
S425
D S1 12.5
S1 12.5
S1 12.5
S2 12.5
S2 12.5
S2 12.5
S3 12.5
S3 12.5
S3 12.5
S4 12.5
S4 12.5
S4 12.5
E S1 6.25
S1 6.25
S1 6.25
S2 6.25
S2 6.25
S2 6.25
S3 6.25
S3 6.25
S3 6.25
S4 6.25
S4 6.25
S4 6.25
F S1 3.13
S1 3.13
S1 3.13
S2 3.13
S2 3.13
S2 3.13
S3 3.13
S3 3.13
S3 3.13
S4 3.13
S4 3.13
S4 3.13
G S1 1.56
S1 1.56
S1 1.56
S2 1.56
S2 1.56
S2 1.56
S3 1.56
S3 1.56
S3 1.56
S4 1.56
S4 1.56
S4 1.56
H S1 0.781
S1 0.781
S1 0.781
S2 0.781
S2 0.781
S2 0.781
S3 0.781
S3 0.781
S3 0.781
S4 0.781
S4 0.781
S4 0.781
I NC NC NC PC PC PC
DNA Extraction with a Twist
Most people have heard of or done the DNA extraction either with a kit (like Carolina’s DNA necklace) or from a kitchen chemistry protocol (using shampoo, salt, and a coffee filter).
When doing the strawberry (or other fruit) extraction I like to use frozen due to when the ice crystals form they act like mini daggers and rupture some of the cells (that is why frozen fruit becomes “mushy” when thawed).
DNA Extraction Notes of Interest
Mine is a twist on the kitchen chemistry version. Instead of using shampoo and water for the
extraction buffer use Mrs. Stewart’s Bluing (can be found in the laundry section as it is used to “Make whites whiter”.
The bluing solution will “stain” the DNA blue making it much easier to see.
When using strawberry or some other fruit DNA is plentiful and easy to see as white fluff in normal protocols but when doing the human cheek extraction the DNA is hard to see due to low quantities so the Mrs. Stewart’s is very helpful.
$366 ~ $7 bin + $20 Blanket = $27
=
=
=
$$$ Money Savers $$$
Money Savers
Incubator- Plastic bin wrapped in an electric blanket with a thermometer (cheap fish tank one) to determine what setting is needed for desired temperature.
If moist heat is needed place a shallow dish of water in bin with plates (maybe put a wire rack on top of a shallow pan or dish to put the plates on).
Make your own media (very easy, made with items found at local grocery).
Don’t have Bunsen burners? Use chaffing dishes. No water bath- Use a pot on a hot plate.
Ask Around
Most science equipment and materials have an expiration date or a calibration date and when these items pass that date in industry (especially ones that require FDA approval) must be discarded.
Contact local science companies, university science departments, and hospital research facilities and state your need for any expired supplies that may be past their expiration date.
Want My Lessons or Materials List?
Go to:
lebraves.net/NSTA
Or:
lebraves.net
Then click on the NSTA tab at the top.
Questions or Comments:
Feedback Very Welcomed! =-)
