Starting your own biolab: Difference between revisions
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=== Thermal cycler === | === 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 relatively 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. Newer ones are based on Peltier-effect blocks. There are also open-source DIY alternatives such as OpenPCR, though second-hand ones are much cheaper. | |||
=== Electrophoresis tank and power supply === | === Electrophoresis tank and power supply === | ||
| Line 19: | Line 20: | ||
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. | 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. | [[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. | ||
=== Visualiser === | === 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. | 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 === | === 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. | 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 === | === 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 | For sterilizing labware and inactivating living material. We started with a [[Autoclave|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 [[Project:Flow_Hood|flow hood]]. | |||
== Reagents and disposables == | == Reagents and disposables == | ||
| Line 50: | Line 51: | ||
Agarose is purified, very fine, high-quality agar. It is used for making agarose “gels” for gel electrophoresis. | 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 === | === Pipette tips === | ||
| Line 63: | Line 68: | ||
* Petri dishes: we use plastic ones between 80 and 140mm. | * Petri dishes: we use plastic ones between 80 and 140mm. | ||
=== Chemicals for inactivation and disposal === | |||
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. | |||
[[Category:Biohacking]] | [[Category:Biohacking]] | ||
Revision as of 21:22, 16 June 2016
Congratulations on your decision to start a biolab! Here the London biohackspace has made a few notes on what you might need when you’re starting out. With these hints your new biolab should give you many years of trouble-free service.
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
- Labcoats
- Glasses or goggles
- Disposable gloves. We use nitrile gloves.
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 relatively 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. Newer ones are based on Peltier-effect blocks. There are also open-source DIY alternatives such as OpenPCR, though second-hand ones are 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.
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.
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 inactivation and disposal
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.
