Difference between revisions of "Starting your own biolab"

From London Hackspace Wiki
Jump to navigation Jump to search
 
(One intermediate revision by the same user not shown)
Line 18: Line 18:
 
* Glasses or goggles
 
* Glasses or goggles
 
* Disposable gloves. We use nitrile gloves. In addition to preventing contamination of your hands with your sample, nitrile gloves can be disinfected to prevent contamination of your sample.
 
* Disposable gloves. We use nitrile gloves. In addition to preventing contamination of your hands with your sample, nitrile gloves can be disinfected to prevent contamination of your sample.
 +
 +
=== Centrifuge ===
 +
Separates immiscible liquids, or separates solid from liquid. We mostly use microcentrifuges, which take PCR tubes and spin at a maximum of about 13000 RPM (though lower speeds, and larger centrifuges, are fine for many protocols if that's all you have).
 +
 +
High-speed centrifuges can be very dangerous and should be treated with respect.
  
 
=== Thermal cycler ===
 
=== Thermal cycler ===
Line 29: Line 34:
  
 
[[Project:Gel_electrophoresis_box|Our first electrophoresis set-up was home-made]], using laser-cut acrylic gel boxes and standard laboratory variable-output power supplies.  
 
[[Project:Gel_electrophoresis_box|Our first electrophoresis set-up was home-made]], using laser-cut acrylic gel boxes and standard laboratory variable-output power supplies.  
 +
 +
We do agarose gel electrophoresis. See below for information on agarose.
  
 
=== Visualiser ===
 
=== Visualiser ===
Line 53: Line 60:
  
 
To prevent contamination of samples. These force air through a HEPA filter onto the sample in a laminar flow, ensuring that the only air touching the sample is filtered. Flow hoods additionaly create a closed cycle of air, preventing aerosolised samples from contaminating the rest of the lab. Our current flow cabinet is home-made. We have plans to turn it into a [[Project:Flow_Hood|flow hood]].
 
To prevent contamination of samples. These force air through a HEPA filter onto the sample in a laminar flow, ensuring that the only air touching the sample is filtered. Flow hoods additionaly create a closed cycle of air, preventing aerosolised samples from contaminating the rest of the lab. Our current flow cabinet is home-made. We have plans to turn it into a [[Project:Flow_Hood|flow hood]].
 +
 +
=== Refrigerator and freezer ===
 +
 +
Storing reagents and bacteria in a fridge or freezer keeps them viable for longer. We currently use standard mini fridges (at 4 C) and mini freezers (at around -20 C). A -80 C freezer is the best option for long-term storage, but these are large and expensive both to buy and to run, so we don't use one.
 +
 +
=== Incubator ===
 +
 +
Incubators maintain a constant temperature and are ideal for growing bacterial colonies (among other things). If you are planning on culturing in liquid media (such as lb broth) you should look for a shaking incubator, which has a shaking platform to keep the liquid moving. If you are culturing in petri dishes shaking is not necessary.
 +
 +
Our first incubator was home-made from an ice box, an Arduino, a temperature sensor, and a heat lamp. Current incubators are donated.
  
 
== Reagents and disposables ==
 
== Reagents and disposables ==
Line 81: Line 98:
  
 
For general lab disposal we have settled on Virkon (in a [https://github.com/london-biohackers/biohackspace-sops/blob/master/lbl04007.rst "kill bin"]). For inactivation of ethidium bromide (or equipment contaminated by it), we use bleach as part of the [https://github.com/london-biohackers/biohackspace-sops/blob/master/lbl09001.rst Armour protocol]. GMOs are autoclaved.
 
For general lab disposal we have settled on Virkon (in a [https://github.com/london-biohackers/biohackspace-sops/blob/master/lbl04007.rst "kill bin"]). For inactivation of ethidium bromide (or equipment contaminated by it), we use bleach as part of the [https://github.com/london-biohackers/biohackspace-sops/blob/master/lbl09001.rst Armour protocol]. GMOs are autoclaved.
 +
 +
== Suppliers ==
 +
 +
Most of the traditional lab suppliers have an application process, and don't ship to residential addresses. This is unfortunate but makes some sense as they sell quite hazardous chemicals.
 +
 +
* [http://www.sigmaaldrich.com/united-kingdom.html Sigma-Aldrich]: Buffer, plastics, agarose, primers, PCR master mix and its consituents
 +
* [http://www.nbsbio.co.uk/ NBS Bio]: Pipette tips, PCR tubes, buffer, ladders, PCR master mix. As you can see, significant overlap with Sigma-Aldrich, but NBS Bio tends to be slightly cheaper.
 +
* [http://ebay.co.uk eBay]: used equipment, plastics
  
 
[[Category:Biohacking]]
 
[[Category:Biohacking]]

Latest revision as of 14:21, 18 June 2016

This page describes the equipment you will need to start up your own microbiology lab.

Although this list focuses on equipment and reagents, there is a lot more to a lab than this. For more information, have a look at:

Essential equipment

This will, of course, vary depending on what you want to do. With its focus on molecular- and microbiology, the London Hackspace acquired the following equipment, roughly in this order:

Personal protective equipment

The purpose of PPE is to prevent harmful substances from contacting your skin or eyes. In addition to lab basics (such as enclosed shoes), we use:

  • Labcoats
  • Glasses or goggles
  • Disposable gloves. We use nitrile gloves. In addition to preventing contamination of your hands with your sample, nitrile gloves can be disinfected to prevent contamination of your sample.

Centrifuge

Separates immiscible liquids, or separates solid from liquid. We mostly use microcentrifuges, which take PCR tubes and spin at a maximum of about 13000 RPM (though lower speeds, and larger centrifuges, are fine for many protocols if that's all you have).

High-speed centrifuges can be very dangerous and should be treated with respect.

Thermal cycler

A thermal cycler (or PCR machine) is generally used to amplify segments of DNA, for example to test for the presence of certain sequences.

Our current thermal cyclers were acquired through donations, but they are available cheaply on eBay. The job of a thermal cycler is to cycle through temperatures ranging between (typically) 4 and 95 degrees C. Older ones are large and bulky and are, essentially, miniature refrigerators stuck to miniature space heaters. Newer ones are based on Peltier-effect blocks. There are also open-source DIY alternatives such as OpenPCR. The open-source options have the advantage of a well-documented protocol for computer control. Second-hand commercial units are, however, much cheaper.

Electrophoresis tank and power supply

Used to move DNA fragments through a substrate at a speed proportional to the length of the DNA. Part of the process of PCR result visualisation.

Our first electrophoresis set-up was home-made, using laser-cut acrylic gel boxes and standard laboratory variable-output power supplies.

We do agarose gel electrophoresis. See below for information on agarose.

Visualiser

Part of the process of PCR result visualisation. A visualiser produces light at a wavelength appropriate to your visualisation agent, making it glow a a visible wavelength. The visualiser you use will depend on agent you use for visualisation. For example, we use ethidium bromide, which has an absorption spectrum centred around 300nm, and thus have a visualiser based around a 300nm ultraviolet light.

Our first visualisation set-up involved a simple 300nm fluourescent tube. Note that ultraviolet light at this frequency (UVB) is carcinogenic! Ensure that your eyes are suitably covered, and no skin is exposed, when using the visualiser.

Visualisers are also available on eBay. Unfortunately 300nm visualisers are rather expensive. You can avoid the expense by using a visualisation agent which uses a different frequency of light, such as sybr-safe -- but the increased cost of sybr-safe will eventually offset this savings.

Pipettes

Perhaps the most-used tool in mol bio. You will need several. We tend to use 10, 20, 50, and 100 ul volume pipettes most often. The standard brand is Gilson; these (and knock-offs) are available on eBay.

Pipettes are non-disposable but use disposable tips, which are made of plastic. Different brands (and different pipette volumes) require different sizes of tip.

Autoclave

For sterilizing labware and inactivating living material. We started with a pressure cooker, but autoclaves often come with useful features, such as a vacuum cycle for more effective sterilisation of porous material.

We currently use a Prestige classic Autoclave

Laminar flow cabinet or hood

To prevent contamination of samples. These force air through a HEPA filter onto the sample in a laminar flow, ensuring that the only air touching the sample is filtered. Flow hoods additionaly create a closed cycle of air, preventing aerosolised samples from contaminating the rest of the lab. Our current flow cabinet is home-made. We have plans to turn it into a flow hood.

Refrigerator and freezer

Storing reagents and bacteria in a fridge or freezer keeps them viable for longer. We currently use standard mini fridges (at 4 C) and mini freezers (at around -20 C). A -80 C freezer is the best option for long-term storage, but these are large and expensive both to buy and to run, so we don't use one.

Incubator

Incubators maintain a constant temperature and are ideal for growing bacterial colonies (among other things). If you are planning on culturing in liquid media (such as lb broth) you should look for a shaking incubator, which has a shaking platform to keep the liquid moving. If you are culturing in petri dishes shaking is not necessary.

Our first incubator was home-made from an ice box, an Arduino, a temperature sensor, and a heat lamp. Current incubators are donated.

Reagents and disposables

Agarose

Agarose is purified, very fine, high-quality agar. It is used for making agarose “gels” for gel electrophoresis.

Autoclave tape

Changes colour when autoclaving was successful (i.e. 15 minutes at 121 degrees C)

Pipette tips

You will go through these very rapidly. Note that pipettes of differing maximum volume tend to require different size tips.

Glasswear

You can't have too little. We use a lot of 500ml beakers. If you will be growing living things, you will want closeable jars.

Plastics

  • Petri dishes: we use plastic ones between 80 and 140mm.

Chemicals for aseptic procedure, inactivation, and disposal

We use 70% ethanol solution as disinfectant.

For general lab disposal we have settled on Virkon (in a "kill bin"). For inactivation of ethidium bromide (or equipment contaminated by it), we use bleach as part of the Armour protocol. GMOs are autoclaved.

Suppliers

Most of the traditional lab suppliers have an application process, and don't ship to residential addresses. This is unfortunate but makes some sense as they sell quite hazardous chemicals.

  • Sigma-Aldrich: Buffer, plastics, agarose, primers, PCR master mix and its consituents
  • NBS Bio: Pipette tips, PCR tubes, buffer, ladders, PCR master mix. As you can see, significant overlap with Sigma-Aldrich, but NBS Bio tends to be slightly cheaper.
  • eBay: used equipment, plastics