LitCovid-sentences  

PMC:7143804 / 10922-13453 JSONTXT 6 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T90 0-4 Sentence denotes 1.3.
T91 5-40 Sentence denotes Multiple Displacement Amplification
T92 41-194 Sentence denotes The proof-of-principle amplification of choice is a MDA reaction, which is a non-specific isothermal method of amplification performed around 30 °C [50].
T93 195-285 Sentence denotes MDA is a method of whole genome amplification (WGA), as it amplifies all present DNA [52].
T94 286-356 Sentence denotes It is commonly used when the initial amount of DNA sample is very low.
T95 357-492 Sentence denotes After the WGA is performed, a sequence specific amplification can be done since the quality the amplified DNA by MDA is very high [53].
T96 493-629 Sentence denotes The amplification reaction is illustrated below in Figure 1 (the contour of the amplified sequence is highlighted in black for clarity).
T97 630-846 Sentence denotes Starting with a double stranded DNA (dsDNA) molecule, a denaturation step at 95 °C is required, giving the random hexamer-primers and the ϕ29 DNA polymerase access to the bases of single stranded DNA (ssDNA) strands.
T98 847-916 Sentence denotes The hexamers anneal themself to aleatory parts of the ssDNA sequence.
T99 917-985 Sentence denotes These hexamers work as initiation sites for the ϕ29 DNA polymerases.
T100 986-1100 Sentence denotes After denaturation at 95 °C, the mixture is cooled down to ice temperature and the rest of the reagents are added.
T101 1101-1301 Sentence denotes The mixture is heated up to ~30 °C so the polymerase starts to complete the complementary ssDNA sequence, creating again a dsDNA strand, eventually it encounters a hexamer from another annealing site.
T102 1302-1439 Sentence denotes Once this happens the polymerase will lift up that hexamer and starts to separate the amplified sequence formed from that annealing site.
T103 1440-1537 Sentence denotes As the polymerase displaces the formed strand ahead of it, it continues to complete the sequence.
T104 1538-1757 Sentence denotes The displaced strand becomes a new ssDNA strand and therefore, it gives new sites for more primers to attach and initiation sites for the polymerase, continuing the amplification, and thus creating a web of DNA strands.
T105 1758-1844 Sentence denotes Finally, the inactivation of the polymerase is done by heating up the system to 65 °C.
T106 1845-2089 Sentence denotes Even though MDA is considered an isothermal process, prior to the reaction and to the addition of most reactants, the dsDNA and a buffer are heated up to 95 °C to denature the dsDNA to ssDNA and to give hexamers the initial access to the ssDNA.
T107 2090-2170 Sentence denotes After the amplification reaction, the polymerase has to be inactivated at 65 °C.
T108 2171-2314 Sentence denotes However, this does not require fast temperature changes, as would be the case with, for example, the temperature cycling in PCR amplifications.
T109 2315-2531 Sentence denotes This, together with the robustness of the amplification (it is a self-limiting reaction that amplifies all present DNA [50]) makes MDA perfectly suitable as proof-of-principle amplification reaction for such devices.