| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T61 |
0-135 |
Sentence |
denotes |
To detect SARS-CoV-2 RNA with RT-LAMP, we used the WarmStart Colorimetric RT-LAMP 2X Master Mix (DNA and RNA) from New England Biolabs. |
| T62 |
136-254 |
Sentence |
denotes |
This mix contains two enzymes, an engineered reverse transcriptase (RTx) and a strand-displacing polymerase (Bst 2.0). |
| T63 |
255-506 |
Sentence |
denotes |
In addition, the reaction mixture contains oligonucleotide-based aptamers that function as reversible temperature-dependent inhibitors, ensuring that the reaction only runs at an elevated temperature (WarmStart) to avoid nonspecific priming reactions. |
| T64 |
507-719 |
Sentence |
denotes |
Several primer sets were recently proposed for RT-LAMP–based detection of SARS-CoV-2 RNA by Zhang et al. (11) and by Yu et al. (10), and these primer sets were subsequently validated with in vitro–translated RNA. |
| T65 |
720-916 |
Sentence |
denotes |
We prepared and tested two primer sets for different RNA sections of the SARS-CoV-2 genome, the N-A set targeting the N gene and the 1a-A set targeting open reading frame (ORF) 1a (table S1) (11). |
| T66 |
917-1117 |
Sentence |
denotes |
Figure 1A shows that the oligonucleotide set for the N gene was capable of detecting 100 IVT RNA molecules in a test reaction with 1 μl of RNA solution, as evidenced by the red-to-yellow color change. |
| T67 |
1118-1170 |
Sentence |
denotes |
The reaction was conducted for up to 1 hour at 65°C. |
| T68 |
1171-1262 |
Sentence |
denotes |
For time points > 30 to 35 min, the negative control frequently became yellowish (Fig. 1A). |
| T69 |
1263-1363 |
Sentence |
denotes |
This was caused by spurious amplification products, which is a well-known problem with RT-LAMP (14). |
| T70 |
1364-1568 |
Sentence |
denotes |
Analysis by gel electrophoresis revealed clearly distinct banding patterns for the correct RT-LAMP reaction products (lanes with ≥100 molecules IVT RNA input) and the spurious reaction products (Fig. 1B). |
