BioLab Manual: Difference between revisions
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= Safety = | = Safety = | ||
All users of the Biolab must follow the [[BioLab Usage Guidelines]] for their own and other user's safety. | |||
= Protocols = | |||
== | |||
Instructions for [[E-coli RFP Transform |(a "Blink LED" for easy first transformations)]] | |||
Instructions for [[Stock Solutions|making general stock solutions]] | Instructions for [[Stock Solutions|making general stock solutions]] | ||
| Line 33: | Line 18: | ||
* [[DNA extraction using Chelex 100]] | * [[DNA extraction using Chelex 100]] | ||
* [[DNA extraction and precipitation with ethanol / isopropanol]] | * [[DNA extraction and precipitation with ethanol / isopropanol]] | ||
* [[DNA extraction and precipitation with phenol-chloroform]] | |||
* [[Make and run an agarose gel]] | * [[Make and run an agarose gel]] | ||
* [[ | * [[Solid Media Bacterial Culture]] | ||
* [[ | * [[Liquid Media Bacterial Culture]] | ||
* [[PCR]] | * [[PCR]] | ||
* [[Miniprep | Anachem Miniprep (plasmid purification)]] | * [[Miniprep | Anachem Miniprep (plasmid purification)]] | ||
* [[LBA]] (LB ampicillin) | |||
We also have [https://github.com/london-biohackers/biohackspace-sops/blob/master/index.rst protocols on our github SOPs page] | |||
== Untested Protocols== | == Untested Protocols== | ||
The protocols we would like to get working in the Biohackspace. | The protocols we would like to get working in the Biohackspace. | ||
== Media Recipes == | == Media Recipes == | ||
| Line 50: | Line 37: | ||
* [[HS Medium]] | * [[HS Medium]] | ||
* [[Kombucha Medium]] | * [[Kombucha Medium]] | ||
* [[TBE]] | |||
= Equipment = | = Equipment = | ||
| Line 76: | Line 64: | ||
== Strains == | == Strains == | ||
* [[E. coli DH5α]] | * [[E. coli DH5α|''E. coli'' DH5α]] | ||
* [[V. fischeri]] | * ''[[V. fischeri]]'' | ||
* [[P. polycephalum]] | * ''[[P. polycephalum]]'' | ||
* ''[[G. hansenii]]'' | |||
* ''[[Dinoflagellate|''P. fusiformis, P. lunula'']] | |||
== Reagents == | == Reagents == | ||
The reagents we currently have stocked | [[:Category:BiolabInventory]] | ||
The reagents we currently have stocked | |||
{| class="wikitable sortable" style="float:left" | {| class="wikitable sortable" style="float:left" | ||
Latest revision as of 19:58, 2 October 2016
Safety
All users of the Biolab must follow the BioLab Usage Guidelines for their own and other user's safety.
Protocols
Instructions for (a "Blink LED" for easy first transformations)
Instructions for making general stock solutions
For more extensive databases of protocols:
Established Protocols
The protocols we have got working in the Biohackspace:
- DNA extraction using Chelex 100
- DNA extraction and precipitation with ethanol / isopropanol
- DNA extraction and precipitation with phenol-chloroform
- Make and run an agarose gel
- Solid Media Bacterial Culture
- Liquid Media Bacterial Culture
- PCR
- Anachem Miniprep (plasmid purification)
- LBA (LB ampicillin)
We also have protocols on our github SOPs page
Untested Protocols
The protocols we would like to get working in the Biohackspace.
Media Recipes
Equipment
To create a page for a piece of lab equipment fill out this form and make sure you put "Biohacking" as the subcategory.
We also have an array of glassware and consumables.
This site has a directory of service engineers for various biomedical/biochemical equipment:
http://www.labwrench.com/?equipment.requestAQuote/categoryNo/707/Mass-Spectrometers/
Strains
Reagents
The reagents we currently have stocked
| Reagent | Brand | Location |
|---|---|---|
| Bacto Agar | Shelves | |
| Bacto Yeast Extract | Shelves | |
| LB broth powder | Shelves | |
| TBE buffer x10 | Shelves | |
| Sodium Acetate | Shelves | |
| Ethidium Bromide | Drawers | |
| Tri-Sodium Citrate-2 Water | Shelves | |
| Magnesium Chloride | Shelves | |
| Calcium Chloride | Shelves | |
| Agarose | Shelves | |
| Glycerin | Shelves | |
| Ampicillin Sodium Salt (powdered) | Fridge | |
| Kanamycin Sulphate (powdered) | Fridge | |
| IPTG | Freezer | |
| Proteinase K 1mg/ml | Freezer |
| Reagent | Description | Brand |
|---|---|---|
| pUC19 control DNA | NEB #N3041A | NEB |
| Q5 Hi-GC enhancer | 5x concentrate | NEB |
| Reagent | Cutting Site | Brand |
|---|
See also: Project:Making our own reagents
link to shopping list G spreadsheet
Suppliers
- Sigma-Aldrich - Good for custom primers
- Cole-Parmer
- Farnell
- Chang bioscience - pipettes
- NBSBio - Good for ladders and cheap agarose (only low concentration) NB: Be sure to navigate to http://www.nbsbio.co.uk NOT http://nbsbio.co.uk, or their payment system doesn't work. Weird. Standard shipping cost is £15
- Web scientific - Taq and ladder
- New England Biolabs They sell very cheap taq. We could also look at their restriction enzymes. However atm they won't sell to us as they only sell to universities or registered biolabs. See this post and this post. Once we have all our registrations for LBL sorted we could try again.
- Thermoscientific - Recommended for good value restriction enzymes. Not tried yet.
- VWR - Big supplier of chemicals, glassware and various lab stuff. Sounds very positive about selling to the hackspace.
- Invitrogen We failed their due diligence check. See post here and here. They have cheap TBE.
- IDT - possible source for primers. Haven't tried them yet.
- cheaplabstuff (ebay) - Cheap 1.5ml and 15ml tubes in small quantities. Also pasteur pipettes.
- microsupplies (ebay) - Cheap 5-200ul and 100-1000ul tips by the 1000
- Cambridge Bioscience - Not yet used, may supply GelRed or GelGreen (enquiry by Bene)
Also see general hackspace suppliers page.
