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| -rw-r--r-- | E.coli Transformation With PEG+MgSO4.md | 34 |
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diff --git a/E.coli Transformation With PEG+MgSO4.md b/E.coli Transformation With PEG+MgSO4.md index 00e968b..b87a859 100644 --- a/E.coli Transformation With PEG+MgSO4.md +++ b/E.coli Transformation With PEG+MgSO4.md @@ -11,33 +11,33 @@ The following protocol is designed to make it as easy for a beginner to make the * Optional centrifuge * Optional autoclave/pressure cooker -## 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. +## 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. * 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 +* 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. * 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 +* 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 -* 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. +* 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. * 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” +* 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”. ## Transforming E.coli by a modified TSS method: |