BISC209: Lab3

Lab 3: Continue Soil Microbial Communities & Diversity Project

Protocol: PCR Amplify Genomic DNA from Universal Bacterial Primers

DNA diversity: complete prep for pyrosequencing? OR rDNA library steps: (Use soil DNA from lab 2) DNA library:

Clean and PCR Amplify for rDNA using Irene's universal primers (Irene's notes)- 3 hours

CULTURES: (Primary plates) observe-count, describe, select soil microbes for further culturing (compare anaerobic, aerobic, microaerophillic) - search web for pics of microbes.

techniques: colony morphology, number (see mIcrobial safari= wagner) and problem solve to "discover" 2 isolation methods (serial dilution and pour plates vs streaking for isolation)

Isolation to pure colony step: #1: How will they pick what/where on plate to isolate? (secondary plates) Each student picks different 4-6 colonies and restreaks on same media (4-6 different plates). Incubate room temp.grow 24 hours, move to CR.

Microscopy introduced: select something that looks isolated and stain and compare to stock cultures.

Control stocks for: gram pos, gram neg and capsule, acid fast and endospore

Simple stain Gram stain

What do you learn from this?