TITLE: Preparation of Binding Buffer (BB)
AUTHORS: Nicola O'Reilly, Svend Kjaer, Maria Greco

[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 FCI 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 binding buffer for the </span><a href="https://www.protocols.io/private/092BEFAA830211EABF5D0242AC110002" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">RNA extraction</span></a><span> protocol.</span></div></div>

[STEPS]
?. Please select between the recipes for 2 L + 80 ml Binding buffer BB, 4 L + 160 ml Binding buffer BB and 5 L + 200 ml Binding buffer BB.
?. Weigh out  and add to a 2 L beaker.
[GuHCl]
?. Add . Stir and heat to  to dissolve.
[freshly drawn milliQ]
60 °C
?. Add , .
[3M sodium acetate]
?. Add . Stir well to mix.
[Tween-20]
?. Vacuum filter into sterile bottles.
?. Label with batch and date.
?. Store Binding Buffer BB buffer in a flammables cupboard at  ( – ).
0 Room temperature
15 °C
25 °C
?. Add  to a 5 L beaker.
[freshly drawn milliQ water]
?. Weigh out  in aliquots using a 3 L beaker and carefully add to 5 L beaker. Stir and heat to  to dissolve. A sterile pipette may be required to stir.
[GuHCl]
60 °C
?. Once dissolved, make up to 4 L with Isopropanol (approx. ) using a 1 L graduated cylinder and transferring buffer to second 5 L beaker.
1600 ml
?. Add , .
[3M sodium acetate]
?. Add . Stir well to mix.
[Tween-20]
?. Vacuum filter into sterile bottles.
?. Label with batch and date.
?. Store Binding Buffer BB buffer in a flammables cupboard at  ( – ).
0 Room temperature
15 °C
25 °C
?. Once dissolved, make up to 2 L with  Isopropanol (approx. ) using a 1 L graduated cylinder.
800 ml
?. Add  to a 5 L beaker.
[freshly drawn milliQ water]
?. Weigh out  in aliquots using a 3 L beaker and carefully add to 5 L beaker. Stir and heat to  to dissolve. A sterile pipette may be required to stir.
[GuHCl]
60 °C
?. Once dissolved, make up to 5 L with Isopropanol (approx. ) using a 1 L graduated cylinder and transferring buffer to second 5 L beaker.
2 L
?. Add , .
[3M sodium acetate]
?. Add . Stir well to mix.
[Tween-20]
?. Vacuum filter into sterile bottles.
?. Label with batch and date.
?. Store Binding Buffer BB buffer in a flammables cupboard at  ( – ).
0 Room temperature
15 °C
25 °C