Description
My Lab Overview: The HHMI Phagehunters laboratory will be very different from the majority of laboratories that you have previously completed in your other college courses. You will be involved in a semester-long authentic biology research project in which you and your lab partner will discover, characterize, and even name an organism currently unknown to science! Your target is a virus, or bacteriophage, that infects a bacterium known as Bacillus thurengensis (BtK), a harmless organism that is related to Bacillus anthracis, the causative agent of anthrax, and Bacillus cereus, a human pathogen. You will learn how to do independent research in this laboratory as you master skills like bacterial culturing, pipetting, sterile technique, electron microscopy, DNA isolation and electrophoresis, and many others. This will not be your typical “cook book” lab where you come in each day and perform a set of actions—instead it will be more akin to a “Choose Your Own Adventure” book, where the decisions that you make one week will dictate your work for the rest of the semester!
My Lab Manual is: HHMI Phagehunters lab manual
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Explanation & Answer
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Lab Section #12
3/30/2016
Lab Report: Methods, Results, and Discussion
Materials and Methods:
For the experiment we first collected a soil sample in plastic bag of at least
0.5g. After obtaining the sample, we began soil enrichment of the soil, using the
protocol of isolating the phage.
Isolate a Nobel Phage from the Environment: Enrichment of Environment
The enrichments were divided into two days. The first day we took 0.5g of the soil
using a spatula and mixed it with 35mL of T-soy broth and 4 mL of the Btk. Then, we
added them all to a 50mL bioreactor. After 24 hours the teaching assistant enhanced
the mixture with additives for better results, and used a microcentrifuge pipette to
dispense some of the liquid in a microcentrifuge tube. On day two of the enrichment,
which occurred the next week, two plates were made. The first plate used Btk and
TA (Top Agar). 4.5mL of TA were poured into 0.5mL of the infected Btk and 10
minutes were waited while it completely solidified. The second plate was prepared
with slices labeled as 100, 10-1, 10-2, 10-3, 10-4, and SM buffer. To make these, 4
microcentrifuge tubes were taken and labeled -1, -2, -3, and -4. The 0 tube contained
the solution from the bioreactor. Extracting the SM buffer with the microcentrifuge
pipette, 90microlite was added to each of the 4 tubes -1, -2, -3, and -4; then
10microliter from the 0 tube were adding to -1 ; 10microliter from -1 to -2; 10
microliter from -2 to -3; and 10 microliter from -3 to -4. For the sixth tube, 1.0mL of
SM buffer were added to make a negative control for it. Meanwhile, TA a...
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