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David Coil edited General notes on molecular and microbiology.md
almost 10 years ago
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#General notes on molecular and microbiology
This
section workflow assumes basic knowledge of molecular biology and sterile technique. The starting point
for this workflow is the collection of microbes from a surface with a cotton swab.
Here we We will cover the steps necessary to take a sample through plating, dilution streaking, overnight growth, creating a glycerol stock, 16s rDNA PCR, and preparation for Sanger sequencing to determine the identity of your bacterial isolate.
Throughout
this the "Isolation" section we refer to "agar" and "culture media". The choice of media will depend on the goals of the particular project. Some factors to consider when selecting media and conditions for growth include:
1. What type of organism do you want to isolate?
...
3. How much time is available for growth and isolation?
+
things many bacteria will grow faster at higher temperatures, but at some point, a threshhold will be reached and many species won't grow
+ you will be able to isolate a greater diversity of organisms if you allow a long time for slow-growing things to grow
5. What types of equipment are available to you?
+ if something grows most happily at \(37\,^{\circ}\mathrm{C}\), then you will need to have an incubator and shaker available at that temperature
For our previous work we have simply used a rich growth media such as lysogeny broth (LB)
and growth at either room temperature or \(37\,^{\circ}\mathrm{C}\).