Author: Kenneth N. Hass; Mengdi Bao; Qian He; Myeongkee Park; Peiwu Qin; Ke Du
Title: Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection (IMPACT) System for Rapid Viral DNA Sensing Document date: 2020_3_20
ID: d840uu3e_24
Snippet: Solid-Phase CRISPR Cas12a Detection: LbCas12a (New England BioLabs, Inc.) with a concentration of 50 nM was pre-assembled with 62.5 nM crRNA (IDT, Inc.) at room temperature for 10 min. Then, LbCas12a-crRNA complexes were mixed with 1 x Binding buffer and 14.75 µl Nuclease Free Water (IDT, Inc.) to reach a 20 ul reaction volume. After adding ASFV target DNA with various concentrations into the mixture, the detection assay was activated and immedi.....
Document: Solid-Phase CRISPR Cas12a Detection: LbCas12a (New England BioLabs, Inc.) with a concentration of 50 nM was pre-assembled with 62.5 nM crRNA (IDT, Inc.) at room temperature for 10 min. Then, LbCas12a-crRNA complexes were mixed with 1 x Binding buffer and 14.75 µl Nuclease Free Water (IDT, Inc.) to reach a 20 ul reaction volume. After adding ASFV target DNA with various concentrations into the mixture, the detection assay was activated and immediately injected into the IMPACT chip. The device was incubated on a hotplate at 37°C for 2 hrs to allow for optimized reaction. Finally, the cleaved product was retrieved from the channel and evaluated by a custom designed fluorometer.
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