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Thomas Joseph


Blog post: The rumors of my death ...
Feb 03, 2012, 2:52pm

Blog post: Wednesday Micro Hits
Aug 17, 2011, 6:00am

Comment on Hello World!
Jul 26, 2011, 9:44am

Comment on Is it worth getting an education?
Jul 08, 2011, 9:04am
Friends (8): See all?

Ash

Prabodh Kandala
Texas Tech University Health Science Center

Brian Krueger, PhD
Columbia University Medical Center
Labs
Primary Lab

The TJ Lab
Classified
User Information
Positon:
Doctor
Homepage:
http://bioen...blogspot.com
Projects:
Greenhouse gas production, microbial communities in agriculture
Interests:
Molecular Biology and Microbiology
Heroes:
Raphael (the TMNT)
Non-Scientific Interests:
Amateur Astronomer, Music (Alt. Rock, Classic Rock, Funk), Reading (Sci-Fi mostly)
Protocols
Aklaline Lysis Plasmid Isolation (High Throughput)
Fri, Sep 03, 2010, 1:14 pm CDT
A quick and dirty protocol that yields double stranded plasmid template of approximately 3 mg per sample. Colonies are grown in 96 deep well plates (~1 mL sample volume), and then subjected to alkaline lysis.
Keywords: Alkaline Lysis, plasmid preparation, high throughput
Bead Beating DNA Extraction Protocol
Fri, Sep 03, 2010, 1:16 pm CDT
Soils can be a pain in the rear to extract DNA from. We've tried to work this protocol to eliminate as much bias as possible from the final prep. Bead beat too long and you'll ruin the DNA from your Gram negatives. Bead beat too short and you risk not cracking open your Gram positives. To increase final DNA yields, you could also try adding 1 to 2 uL of GlycoBlue with your 95% EtOH precipitation.
Keywords: bacterial DNA isolation, bead beating
EtOH Precipitation
Fri, Sep 03, 2010, 1:07 pm CDT
The following PDF is an article that was published by the now defunct Bethesda Research Laboratories back in 1985 (before half the people who read this were even born). It discusses the effects of both incubation time and temperature, as well as centrifugation time. The moral is ... spin your DNA prep down for at least 30 minutes. **I don't think posting this poses any copyright issues. My apologies if it does.**
Keywords: EtOH precipitation
Acetone/EtOH Bacterial RNA Isolation
Fri, Sep 03, 2010, 1:15 pm CDT
A protocol used in our lab for isolation of RNA from bacterial cultures. This protocol works equally well for Gram positive and Gram negative bacteria (we've done it with both). The acetone/ethanol mixture is great for long term storage of samples prior to RNA isolation. Cells are theoretically permeabilized and proteins (including RNAses) are inactivated.
Keywords: RNA isolation, RNA storage
Friends