Aug 31, 2020

Public workspacemicrofluidic sars cov 2 lamp protocol

DOI
dx.doi.org/10.17504/protocols.io.bkjjkukn
  • 1IMT
  • Coronavirus Method Development Community
  • covid19-test
Alexandru Salceanu
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DOI: dx.doi.org/10.17504/protocols.io.bkjjkukn
Protocol Citation: Monica Simion, Alexandru Salceanu 2020. microfluidic sars cov 2 lamp protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bkjjkukn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Tested Protocol for SARS CoV-2 detection using LAMP Amplification
Created: August 31, 2020
Last Modified: September 01, 2020
Protocol Integer ID: 41291
Materials
STEP MATERIALS
Reagent1L TE Buffer [1X], pH 8.0, Low EDTA (Tris-EDTA; 10mM Tris base, 0.1mM EDTA)G-BiosciencesCatalog #786-152
ReagentWarmStart Colorimetric LAMP 2X Master Mix (DNA and RNA) - 100 rxnsNew England BiolabsCatalog #M1800S
ReagentLAMP Primer MIX N-gene SARS-Cov-2eurofins genomicsCatalog #NA
ReagentWater, nuclease-freeThermo FisherCatalog #R0581
Protocol materials
Reagent1L TE Buffer [1X], pH 8.0, Low EDTA (Tris-EDTA; 10mM Tris base, 0.1mM EDTA)G-BiosciencesCatalog #786-152
ReagentWarmStart Colorimetric LAMP 2X Master Mix (DNA and RNA) - 100 rxnsNew England BiolabsCatalog #M1800S
ReagentLAMP Primer MIX N-gene SARS-Cov-2eurofins genomicsCatalog #NA
ReagentWater, nuclease-freeThermo FisherCatalog #R0581
ReagentLAMP Primer MIX N-gene SARS-Cov-2eurofins genomicsCatalog #NA
ReagentWater, nuclease-freeThermo FisherCatalog #R0581
ReagentWarmStart Colorimetric LAMP 2X Master Mix (DNA and RNA) - 100 rxnsNew England BiolabsCatalog #M1800S
Reagent1L TE Buffer [1X], pH 8.0, Low EDTA (Tris-EDTA; 10mM Tris base, 0.1mM EDTA)G-BiosciencesCatalog #786-152
LAMP Amplification
LAMP Amplification
40m
40m
MIX saliva in 1x TE buffer

Add Amount500 µL of
Reagent1L TE Buffer [1X], pH 8.0, Low EDTA (Tris-EDTA; 10mM Tris base, 0.1mM EDTA)G-BiosciencesCatalog #786-152

and mix with Amount500 µL saliva sample

Incubate at Temperature65 °C for Duration00:10:00
10m

LAMP Colorimeteric Amplification

Add Amount12.5 µL of
ReagentWarmStart Colorimetric LAMP 2X Master Mix (DNA and RNA) - 100 rxnsNew England BiolabsCatalog #M1800S
Add Amount2.5 µL of
ReagentLAMP Primer MIX N-gene SARS-Cov-2eurofins genomicsCatalog #NA
Add Amount8 µL of

ReagentWater, nuclease-freeThermo FisherCatalog #R0581
Add Amount2 µL of product from step 1

Incubate for Duration00:30:00 at Temperature65 °C

Read the colour of the reaction mix

Expected result
Red - SARS Cov 2 - negative
Yellow- SARS Cov2 - positive


30m