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Wanted: the Roche lysis buffer formula

Wanted: the Roche lysis buffer formula
27 March 2020 - Trade secrets

Pharmaceutical company Roche is taking some heat in The Netherlands after a publication by investigative platform "Follow The Money" (Link). Roche has a market share of 80% for the equipment currently used in corovirus testing (real-time PCR) and the hospitals and laboratories that use this equipment complain that Roche is unable to supply key reagents (Link, Link). Problem is that the disposables, consumables (multiwell plates, tube strips) reagents and particularly a buffer solution are tied to the supplier. If you buy the instrument from Roche you rely on them for everything else, just like a printer where ink or toner is also from the same manufacturer.

The lysis buffer is in particular demand but Roche states it is doing the best it can. At the same time, according to the laboratories, the company is also unwilling to share the recipe for the buffer solution. This recipe is a trade secret. Not true, counters Roche, you can find buffer recipes on the internet but then the laboratories complain that their homemade formulations do not really work in Roche equipment.

The issue has been debated in the The Netherlands parliament. If need be, the government should force cooperation from Roche by inventing an emergency law (link). Things are turning nasty: this morning a The Netherlands Roche facility was vandalized with "how many dead?" in sprayed graffiti. The government has asked Former DSM CEO Feike Sijbesma to look into corona testing procedures and into upscaling testing capabilities.

In the meanwhile what is this crucial lysis buffer? It breaks open cells and liberates the biomolecules such as DNA or RNA. I am going to assume here that the Roche machine is a Magna Pure 96, a "high-throughput robotic workstation for fully automated purification of nucleic acids from up to 96 samples for in vitro diagnostic use" (Link). I cannot be sure but the FDA has this page that lists the Roche MagNA Pure LC as a certified extraction tool for use in corona virus testing (real time PRC), to use with the Total NA External_lysis protocol. Then the buffer is a "MagNA Pure 96 External Lysis Buffer". Then the material datasheet lists the following compounds: guanidinium thiocyanate (30-50%), a chaotropic agent and alpha-(4-(1,1,3,3-Tetramethylbutyl)phenyl)-omega-hydroxypoly(oxy-1,2-ethanediyl) (20-25%) which is a surfactant very much like Triton X-100 (link). A datasheet is not a recipe but literature on lysis buffer formulations is impossible to find (Roche?) as it turns out. One 1990 research article here is close: dissolve 120 g of guanidinium thiocyanate in 100 mL 0.1 M Tris hydrochloride (a buffer solution), add 22 mL 0.2 M EDTA, adjust with sodium hydroxide to pH 8 and then add 2.6 g of Triton X-100.

Update! Just in! Roche will share the formula after all. The 1990 publication mentioned above WAS the "you can find the recipe on the internet" Roche recipe (Link). Dutch news organisation RTL Nieuws talked to two of the authors of the 1990 publication who also happen to be Dutch and they explain that the recipe has been refined since and they are skeptical when it comes to the "Game of Words" played by Roche. To paraphrase, "they are in it for the money".

Update 06 April 2020: New research just in with up date 2020 recipe (Link) mentioned by Paul Cotter

Update 17 April: More home brewing here, Charles Swanton of The Francis Crick Institute.