Janet B. Matsen:Stuff I Learned While Picking Up Biology
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This collection is mostly from advice given to me by The Amazing Justin Siegel.
- purifying DNA on a gel column has ~ 90% loss. purifying DNA on a column (non-gel) has ~ 90% recovery. avoid gel purification. (Justin)
- Freeze 'N Squeeze Gel extraction...?
- Purify gels on gel-specific kits. Purify PCR products on PCR product specific kits. The former does a less effective job removing primer dimers. (Justin)
- TB gives you more cells/plasmids than LB
- Use TB (1-2 mL) overnight for plasmid preps (for pSB1A3, pSB3K3)
- Don't transform plates at room temp over the weekend, especially if you rely on Amp resistance. Often you see a lot of contamination! (Janet)
- Don't be surprised if you get contamination on plates with multiple antibiotic resistances. There are plenty of plasmids floating around natural systems with multiple on them already. Bacteria make antibiotics in natural systems to add competition to an an environment. (Mary Lidstrom)