Difference between revisions of "Davidson Missouri W/Davidson Protocols"

Revision as of 21:21, 13 March 2010

General Lab Information

1. How to Keep a Lab Notebook
2. Common molecular reagents
3. Standard Assembly
4. BioBrick Ends
5. Compatibility of Plasmids
6. Ethanol Precipitate DNA (short protocol)
7. glycerolstocks How to Make Glycerol Stocks of Bacteria

Gel Electrophoresis and Purification

1. Pouring an agarose gel
2. Calculate MWs
3. 1kb MW markers
4. Qiagen QIAquick Gel Purification
5. Qiagen QIAquick Column Regeneration Protocol
6. ElectroElute Gel Purification
7. Ligation Protocol

Digestion, Ligation, Transformation

1. Digest DNA with restriction enzymes
2. Double Digest Guide
3. Shrimp Alkaline Phosphatase
4. Choices for Transformation: Heat Shock vs. Zyppy
5. Heat Shock Transformation OR Short version of Heat Shock
6. Zippy Transformation
7. TSS Competent Cell Transformation

Minipreps

1. Choices for Mini-Preps: Promega vs. Zyppy
2. Promega miniprep
3. Zippy Miniprep

Making New Parts and PCR

1. Building dsDNA with Oligos
2. Setting up PCR mixtures
3. PCR and Mg²⁺ concentration
4. Making dsDNA Using Primer Dimers
5. Clean and Concentrate DNA (after PCR, before digestion)
6. Colony PCR to Screen for Successful Ligations

Expression of Phenotypes

1. Using degradation tags on proteins such as GFP
2. Genomic Insertion Protocol
3. 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.
4. When inducing with Arabinose, use "2 µL of stock" (10% w/v L-Arabinose) "to every 1 mL" of LB or other liquid.

Computer Tools We Use

1. All Sites In One Place
2. Optimize your Gel
3. Gene Splitting Web Site
4. PCR Primers w/ BioBricks
5. Promega T_m Calculator
6. List of auto-inducers and their catalog numbers
7. Sequencing at Agencourt Bioscience
8. Sequencing at CUGI
9. Analyzing Sequences with aPe
10. Lance-olator Oligos for dsDNA assembly
11. Access the GCAT-alog of Davidson and MWSU DNA Freezer Stocks
12. Using Apes (A Plasmid Editor)