Setting up a LAMP station can take some work when you are starting from nothing. There is also a lot of choice involved, since our protocols can be adopted in high-throughput labs measuring thousands of samples per day, or on a mountain hut to test friends and family. That’s why we set up a shopping list for you. All the consumables and devices you will or might need are on one page, including what to look for when choosing from different suppliers and substitutes, so you can get by when something goes out of stock.
A staple of any lab, microtubes are essential in a lot of processes. With the usual volumes of 1.5 ml or 2 ml, they are used for mixing reagents together, performing reactions in large volumes, storing solutions and many more things. It's always good to keep a bag at hand. When shopping for microtubes for RT-LAMP, you are mostly free to choose whatever is available to you, but we recommend "low bind" tubes, that are made from a special kind of plastic that keeps the issue of samples sticking to the walls of the tubes to a minimum.
Combining 8 tubes of 200 microlitres each on one strip, PCR strips are an ideal choice for RT-LAMP and bead-LAMP when you have just a few samples to test. They make the colours really pop. When buying PCR strips, make sure to also buy a package of PCR strip caps, this way you can safely seal the reactions. The caps can be domed or flat, if you aren't using a heated lid in your heating instrument, you can go ahead with the domed ones. If you want to do the readout of the LAMP reaction in a qPCR cycler, it's best to use strips that are designed to be optically transparent for qPCR.
For carrying our LAMP reactions on a larger scale, one wants to use these convenient 96-well plates. Each of the wells can hold one RT-LAMP reaction, and wells are spaced so that the plate itself can easily be pipetted with the use of a multichannel pipette to make the work less strenuous. When picking a 96-well plate, think about what you will use to incubate your reactions and make sure your plates and heat source are compatible. If in doubt, contact the manufacturer.
Pipette tips are another one of the essentials. A pipette tip is the single use, small piece of plastic that sits on the bottom part of a pipette and transports the liquid from one solution to another. Filter tips are needed for settings where contamination is a concern, although they are more expensive. We recommend buying boxes filled with tips for convenience.
Falcon tubes are 15 or 50 ml plastic tubes with a screw top, usually graduated. They are convenient for mixing large amounts of solutions, such as master mixes, buffers, beads, and so on. They are good to have on hand for almost everything.
When using 96-well plates, you need to seal the plate before starting the reaction. Plastic adhesive seals are used for this purpose. Make sure to buy these whenever you use 96-well plates. A good seal is crucial to prevent cross contamination, so buy from reputable sources and always seal the plate very well using a plate roller or a plate sealer.
Plastic reagent reservoirs, or troughs, enable the user to pipette one solution with multichannel pipettes at once. Clear, polystyrene or polypropylene material is single use and comes in sterile packaging. It is recommended for use with multichannel pipettes and thus for large sample numbers, and in high-throughput settings.
For handling reactions and inactivated samples, gloves are still a must. RT-LAMP is a simple reaction that can be carried out in low-tech settings, but you must still remember that it is very sensitive and detects up to tens of molecules of DNA/RNA in a reaction. Gloves help to prevent contamination and protect you, your coworkers and your workspace.
To transfer liquids, a micropipette is needed. Variable volume, single channel pipettes are a necessity for any testing scenario. These pipettes can transfer only a specific range of volumes, so make sure you buy enough pipettes to cover the 0.5 - 1000 microlitre range. Make sure that the pipette does not need a specific type of pipette tip. When in doubt, contact the manufacturer.
Multichannels are capable of pipetting 8 or 12 samples at once, significantly increasing the speed of your work. They are always a welcome help when dealing with a larger number of samples. We strongly recommend using them for work with 96-well plates and especially for performing bead-LAMP on a large number of samples.
The magic of bead-LAMP comes from concentrating the sample input RNA thanks to magnetic bead separation. For small numbers of samples, we recommend working using a magnet made for strips. This magnet fits two or four PCR strips and is very convenient to work with. Magnets don't differ much from manufacturer to manufacturer, make sure to get one from a reputable source and that it's compatible with PCR strips.
For bead-LAMP on a large number of samples, you want to work on 96-well plates as you would with regular LAMP. Magnets are made especially for plate separation, and come in many configurations. We find the plate magnets that have the magnetic parts around the perimeter of the well itself to be the most convenient to work with for bead-LAMP, as they enable you to take off all the ethanol from the bottom of the well.
When using 96-well plates and plate seals, a proper sealing of the plate is essential to prevent cross-contamination. For the best seal, a plate roller is recommended. It's a simple roller with a soft, plastic rolling part. For low tech setting, a plate roller can be creatively substituted by any number of household items, just make sure that the seals fit very well on the plate itself. It always pays off to be careful and adhere to measures that prevent contamination.
Tube racks come in all shapes, sizes, and colours. You never want your tubes just laying on the workbench, so stay organised with tube racks. There are many options to choose from and it's mostly up to your personal preference.
These boxes are nothing special, usually paper or plastic and with inserts to fit a number of 1.5 ml tubes comfortably in an organised manner. Boxes help you stay organised and conserve limited storage space in fridges and freezers.
Reliable and precise as any lab instrument should be, a thermocycler is the most precise form of temperature control that you can choose for RT-LAMP reactions. You can run reactions in a standard thermocycler for temperature control or a qPCR thermocycler for simultaneous real-time fluorescent readout of the reaction. The use of a thermocycler may seem like an overkill, but even for low-resource environments it may be a good choice. Many laboratories are getting rid of their old thermocyclers for a bargain, so it is not unusual to find a good deal on a used themocycler for a few hundred dollars. A new thermocycler's cost lies in the range of thousands of dollars.
Considerably cheaper than a thermocycler, a heat block is a device that does just what RT-LAMP needs - a constant temperature. Heat blocks are cheaper than cyclers, and are an ideal choice for RT-LAMP. When shopping for a heat block, make sure it includes an insert for 96-well plates. Our findings suggest that reactions perform the best in heat blocks, and a heated lid is optional for our reactions.
RT-LAMP reactions can be successfully performed using water baths set at 63 degrees. The easiest is the use of a sous-vide cooker, as has been demonstrated in the preprint of our method on bioRxiv. This enables the set-up of RT-LAMP in a truly low-tech manner, and for home testing purposes.
An open source device that incubates samples at 63 degrees and reads out fluorescence in real time. Developed by Andrew Straw and his team at the University of Freiburg, Germany.