Difference between revisions of "Davidson Missouri W/Davidson Protocols"
From GcatWiki
Macampbell (talk | contribs) |
Macampbell (talk | contribs) |
||
| Line 45: | Line 45: | ||
# When inducing with IPTG, use '''3 µL of stock''' (0.2 g/mL = 20% w/v) '''to every 1 mL''' of LB or other liquid. | # When inducing with IPTG, use '''3 µL of stock''' (0.2 g/mL = 20% w/v) '''to every 1 mL''' of LB or other liquid. | ||
# When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid. | # When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid. | ||
| + | # [http://partsregistry.org/AHL List of auto-inducers and their catalog numbers] | ||
| + | |||
'''G. Computer Tools We Use''' | '''G. Computer Tools We Use''' | ||
| Line 52: | Line 54: | ||
# [http://gcat.davidson.edu/iGEM08/bbprimer.html PCR Primers w/ BioBricks] | # [http://gcat.davidson.edu/iGEM08/bbprimer.html PCR Primers w/ BioBricks] | ||
# [http://www.promega.com/biomath/calc11.htm Promega T<sub>m</sub> Calculator] | # [http://www.promega.com/biomath/calc11.htm Promega T<sub>m</sub> Calculator] | ||
| − | # [http:// | + | # [http://gcat.davidson.edu/IGEM06/oligo.html Lance-olator Oligos for dsDNA assembly] |
| + | # [http://gcat.davidson.edu/GCATalog Access the GCAT-alog of Davidson and MWSU DNA Freezer Stocks] | ||
# [[Sequencing at Agencourt| Sequencing at Agencourt Bioscience]] | # [[Sequencing at Agencourt| Sequencing at Agencourt Bioscience]] | ||
# [[Davidson Missouri W/CUGI_Seuqencing| Sequencing at CUGI]] | # [[Davidson Missouri W/CUGI_Seuqencing| Sequencing at CUGI]] | ||
# [http://gcat.davidson.edu/GcatWiki/images/3/3b/Ape_protocol.pdf Analyzing Sequences with aPe] | # [http://gcat.davidson.edu/GcatWiki/images/3/3b/Ape_protocol.pdf Analyzing Sequences with aPe] | ||
| − | |||
| − | |||
# [[Using Apes (A Plasmid Editor)]] | # [[Using Apes (A Plasmid Editor)]] | ||
Revision as of 21:42, 13 March 2010
A. General Lab Information
- How to Keep a Lab Notebook
- Common molecular reagents
- Standard Assembly
- BioBrick Ends
- Compatibility of Plasmids
- Ethanol Precipitate DNA (short protocol)
- glycerolstocks How to Make Glycerol Stocks of Bacteria
B. Gel Electrophoresis and Purification
- Pouring an agarose gel
- Calculate MWs
- 1kb MW markers
- Macherey-Nagel Gel Purification (improved 260/230 ratios)l
- Qiagen QIAquick Gel Purification
- Qiagen QIAquick Column Regeneration Protocol
- ElectroElute Gel Purification
C. Digestion, Ligation, Transformation
- Digest DNA with restriction enzymes
- Double Digest Guide
- Shrimp Alkaline Phosphatase
- Ligation Protocol
- Choices for Transformation: Heat Shock vs. Zyppy
- Heat Shock Transformation OR Short version of Heat Shock
- Zippy Transformation
- TSS Competent Cell Transformation
D. Minipreps
E. Making New Parts and PCR
- Building dsDNA with Oligos
- Setting up PCR mixtures
- PCR and Mg2+ concentration
- Making dsDNA Using Primer Dimers
- Clean and Concentrate DNA (after PCR, before digestion)
- Colony PCR to Screen for Successful Ligations
F. Expression of Phenotypes
- Using degradation tags on proteins such as GFP
- Genomic Insertion Protocol
- When inducing with IPTG, use 3 µL of stock (0.2 g/mL = 20% w/v) to every 1 mL of LB or other liquid.
- When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid.
- List of auto-inducers and their catalog numbers
G. Computer Tools We Use
- All Sites In One Place
- Optimize your Gel
- Gene Splitting Web Site
- PCR Primers w/ BioBricks
- Promega Tm Calculator
- Lance-olator Oligos for dsDNA assembly
- Access the GCAT-alog of Davidson and MWSU DNA Freezer Stocks
- Sequencing at Agencourt Bioscience
- Sequencing at CUGI
- Analyzing Sequences with aPe
- Using Apes (A Plasmid Editor)
