TITLE: Preparation of 0.2M EDTA, pH 8.0
AUTHORS: Nicola O'Reilly

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purpose of examination / Clinical relevance</span></div><div class = "text-block">At the end of 2019, several pneumonia cases were reported in Wuhan, China and the pathogen was confirmed as a new viral strain. World Health organization has named the newly identified coronavirus as 2019-nCoV, also known as COVID19. The disease developed into a dangerous pandemic, posing major challenges to the NHS. Although more research is necessary to better understand the virus, in response to the emergency, simple and rapid testing is essential to identify the virus in infected individuals. This will aid the implementation of efficient interventions to contain the spread, and distinguish healthcare workers who have been infected, and are required to self-isolate, from those showing similar symptoms but which are not 2019-nCoV associated. The latter category may continue to work, alleviating stress on hard-pressed healthcare resources. 2019-nCoV is an RNA virus, and the diagnostic tests detect viral RNA in swabs from patient airways using a reverse transcriptase PCR assay. Samples are submitted to HSL, an accredited reporting laboratory, and transferred to the Crick for testing.</div><div class = "text-block"><span style = "font-weight:bold;">Principles of Examination</span></div><div class = "text-block"><span>This procedure involves the preparation of 0.2M EDTA pH 8.0 for making </span><a href="https://www.protocols.io/private/D1949D20846811EA9B0C0242AC110002" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">L6 virus inactivation buffer</span></a><span style = ":;">.</span></div></div>

[STEPS]
?. Please select between the recipes for 1 L 0.2 M EDTA pH 8.0 and 2 L 0.2 M EDTA pH 8.0.
Work in fume hood!
?. Weigh out , dihydrate and add to a 1 L beaker.
[EDTA disodium salt]
?. Measure out  and add to the beaker.
[MilliQ water]
?. Add a magnetic flea and place on a magnetic stirring plate to mix the solution. The EDTA salt will not go into solution until the pH reaches .
?. Add a newly calibrated pH meter into the solution to observe the pH.
?. To dissolve the salt, add sodium hydroxide (NaOH) pellets to the solution. Add a few pellets at a time and wait until the pellets have fully dissolved before adding more. It may take around  before the pH is at .
[NaOH pellets]
?. Fine tune pH with .
[NaOH]
?. Once fully dissolved (this will take some time so be patient), transfer to 1 L beaker and top up the solution to 1 L using MilliQ water, if necessary.
?. Vacuum filter into sterile bottles.
?. Label with batch and date.
?. Store  at  ( – ).
[EDTA pH 8.0 solution]
0 Room temperature
15 °C
25 °C
?. Weigh out , dihydrate and add to a 2 L beaker.
[EDTA disodium salt]
?. Measure out  and add to the beaker.
[MilliQ water]
?. Add a magnetic flea and place on a magnetic stirring plate to mix the solution. The EDTA salt will not go into solution until the pH reaches .
?. Add a newly calibrated pH meter into the solution to observe the pH.
?. To dissolve the salt, add sodium hydroxide (NaOH) pellets to the solution. Add a few pellets at a time and wait until the pellets have fully dissolved before adding more. It may take around  before the pH is at .
[NaOH pellets]
?. Fine tune pH with .
[NaOH]
?. Once fully dissolved (this will take some time so be patient), transfer to 2 L beaker and top up the solution to 2 L using MilliQ water, if necessary.
?. Vacuum filter into sterile bottles.
?. Label with batch and date.
?. Store  at  ( – ).
[EDTA pH 8.0 solution]
0 Room temperature
15 °C
25 °C