Difference between revisions of "Davidson Protocols"

Revision as of 18:48, 23 September 2013

A. General Lab Information

B. Gel Electrophoresis and Purification

C. Digestion, Ligation, Transformation

D. Minipreps

E. Making New Parts and PCR

F. Expression of Phenotypes

Using degradation tags on proteins such as GFP
Genomic Insertion Protocol
M9CA media (J864-100G from Amresco) + 2mM MgSO₄ (2mL/L of 1 M MgSO₄) + 0.1 mM CaCl₂ (0.1 mL/L 1 M CaCl₂) and optional 0.2% glucose (10 mL/L 20% stock solution).
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 3OC6 (HSL), use a 2000 fold dilution of a 10 mg/mL stock solution. We have dissolved in EtOH which is not the best - degrades with time. Keep this cold.
List of auto-inducers and their catalog numbers
Synergy Machine Protocol

G. Golden Gate Shuffling

H. Computer Tools We Use

General Lab Resources

Discovering New Promoters with Synthetic Biology

TAS2R38 Allele Testing

Evolution of Antibiotic Resistance

@@ Line 53: / Line 53: @@
 # [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC106306/pdf/am002240.pdf Using degradation tags on proteins such as GFP]
 # [[Genomic Insertion Protocol|Genomic Insertion Protocol]]
-# M9CA media (J864-100G from Amresco) + '''2mM MgSO<sub>4</sub>''' (2mL/L of 1 M MgSO<sub>4</sub>) + '''0.1 mM CaCl<sub>2</sub>''' (0.1 mL/L 1 M CaCl<sub>2</sub>) and optional 0.2% glucose (10 mL/L 20% stock solution).
+# '''M9CA media''' (J864-100G from Amresco) + '''2mM MgSO<sub>4</sub>''' (2mL/L of 1 M MgSO<sub>4</sub>) + '''0.1 mM CaCl<sub>2</sub>''' (0.1 mL/L 1 M CaCl<sub>2</sub>) and optional 0.2% glucose (10 mL/L 20% stock solution).
 # 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.