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| author | Cathal Garvey <cathalgarvey@gmail.com> | 2012-09-06 17:03:20 +0100 |
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| committer | Cathal Garvey <cathalgarvey@gmail.com> | 2012-09-06 17:03:20 +0100 |
| commit | 9cbc1945da4b78de153797cb9e7a9a4f2a53f640 (patch) | |
| tree | 2e88661cdfebf34d3cbb16b2868f19bbcab98d45 | |
| parent | b4bd7f8af9e65cb52b92fd27f75a1915a41d9cb9 (diff) | |
| download | cathal-biohacking-protocols-9cbc1945da4b78de153797cb9e7a9a4f2a53f640.tar.gz cathal-biohacking-protocols-9cbc1945da4b78de153797cb9e7a9a4f2a53f640.zip | |
Yet more formatting. I thought github-flavoured-markdown liked line-endings..
| -rw-r--r-- | E.coli Transformation With PEG+MgSO4.md | 30 |
1 files changed, 16 insertions, 14 deletions
diff --git a/E.coli Transformation With PEG+MgSO4.md b/E.coli Transformation With PEG+MgSO4.md index 8caaa06..00e968b 100644 --- a/E.coli Transformation With PEG+MgSO4.md +++ b/E.coli Transformation With PEG+MgSO4.md @@ -13,30 +13,32 @@ The following protocol is designed to make it as easy for a beginner to make the ## Minimum equipment; * Sterile Glass or Pasteur Pipettes or glass droppers, preferably graduated. - * If using glass pipettes, you'll also need a pipette pump and a pipette brush, available cheaply on ebay. Glass droppers must be sterilisable by pressure cooker. Disposible pasteur pipettes should be graduated and sterile, pre-wrapped. + - If using glass pipettes, you'll also need a pipette pump and a pipette brush, available cheaply on ebay. + - Glass droppers must be sterilisable by pressure cooker. + - Disposible pasteur pipettes should be graduated and sterile, pre-wrapped. * 30C incubator - achievable with a pet terrarium thermostat and heat-mat in a polystyrene box. * Crushed ice * Microwave for sterilising media (pressure cooker preferred!) * Sterile work area; - * Bunsen burner or blowtorch-facing-up, or - * Jury-rigged glove-box with inner surface sterilised, or - * A good HEPA-rated air purifier with a fresh filter pointing at work area. + - Bunsen burner or blowtorch-facing-up, or + - Jury-rigged glove-box with inner surface sterilised, or + - A good HEPA-rated air purifier with a fresh filter pointing at work area. * Lab strain, plasmid-free E.coli * Desired DNA plasmid * Ingredients for basic broth: - * Soy Protein (or lactose-free casein) - * Yeast Extract with no added salt (or, balance salt content against your added salt below) - * Salt, non-iodised, preferably without preservatives - * Bromelain or Papain digestive tablets, or Meat tenderiser + - Soy Protein (or lactose-free casein) + - Yeast Extract with no added salt (or, balance salt content against your added salt below) + - Salt, non-iodised, preferably without preservatives + - Bromelain or Papain digestive tablets, or Meat tenderiser * Additional ingredients for TSS-broth: - * PEG-3350, sold under a number of brand names as a laxative: Miralax, GoLytely, GlycoLax, etc. Probably available through pharmacy over the counter, or internet. - * MgSO4, sold as “Epsom Salt” in pharmacies as a bath salt. + - PEG-3350, sold under a number of brand names as a laxative: Miralax, GoLytely, GlycoLax, etc. Probably available through pharmacy over the counter, or internet. + - MgSO4, sold as “Epsom Salt” in pharmacies as a bath salt. * Fine Sieve, Coffee filters, funnel, cotton wool to clarify broth * Glassware - * Preferably actual lab glassware, available from brouwland.com or various ebay sellers. Alternatively; - * Recycled jam jars for “Flasks” - * Smaller jars for “test tubes” - * Wide jars or glass ramekins for “Petri Dishes”. + - Preferably actual lab glassware, available from brouwland.com or various ebay sellers. Alternatively; + - Recycled jam jars for “Flasks” + - Smaller jars for “test tubes” + - Wide jars or glass ramekins for “Petri Dishes”. ## Transforming E.coli by a modified TSS method: 1. Prepare nutrient broth for E.coli (1), and set some aside to make TSS-broth (2). |