Getting Started at the Bench
So you’ve got your keys, your pipettors and are ready to do some good science. Now what? You’ll need to get up-to-speed with some common biochemistry protocols. Here are some resources. Usually, googling what you’re trying to do will lead to additional information.
Contents
Section titled “Contents”1. Protocol Lab Book
Section titled “1. Protocol Lab Book”Molecular Cloning book is awesome and routinely referenced in many labs. Most protocols have a great introduction explaining the fundamentals.
2. Protocols Online
Section titled “2. Protocols Online”- CSH Protocols is the online version of the Lab manual. There are significantly more protocols available, but its not as convenient to browse as a book. Here is a sample of protocols we will use routinely:
- Current Protocols in Molecular Biology is another excellent source for general biochemistry protocols. The chapters below are a very useful general reference:
- Introduction to E. coli, plasmids, etc
- Take a look at Chapters 1, 2, 3 to get started
- OpenWetWare is a Wikipedia-style collection of protocols from different labs from across the world. This site is great for looking at variations in a given protocol from lab to lab.
3. Catalogs and Kits
Section titled “3. Catalogs and Kits”Various molecular biology kits frequently come with a manual that explains the general principles in great detail. Some examples:
- Protein Purification booklets.
4. Useful Buffers for your Bench
Section titled “4. Useful Buffers for your Bench”| Buffer | Recommended Volume | Recipe/Notes |
|---|---|---|
| 1M Tris-Cl pH 8.0 | 1L | Dissolve Tris base in ~700ml H2O. Use conc. HCl to titrate to desired pH. |
| 1M Tris pH 6.8 | 1L | See above |
| 1.5M Tris pH 8.8 | 1L | See above |
| 1M MgCl2 | 0.5 L | |
| 5M NaCl | 0.5 L | |
| 0.5M EDTA pH 8.0 | 0.5L | Titrate to pH 8 with 10N NaOH. EDTA will only dissolve completely at pH 8 |
| TE buffer | 0.5L | 10 mM Tris pH 8, 1 mM EDTA |
| 10% SDS (w/v) | 200 mL | Dissolve SDS in water |
| 10N NaOH | 100 mL | Use a plastic bottle! Glass bottles tend to leach Si into concentrated NaOH. This may cause the lid to get stuck, break the bottle over time and leach unwanted byproducts into your solutions. The reaction is highly exothermic, add very slowly and keep everything on ice. |
| 10X PBS | 1L | |
| 1% agarose | 250 mL | Dissolve in 1x TAE buffer. Make sure agarose does not boil over when you melt in microwave |
| Miniprep buffer I | 250 mL | store at 4C |
| Miniprep buffer III | 250 mL | store at 4C |