Journal: Science Advances

Article Title: Scalable acoustic virtual stirrer for enhanced interfacial enzymatic nucleic acid reactions

doi: 10.1126/sciadv.adt6955

Figure Lengend Snippet: ( A ) Schematic diagrams depicting the polymerization catalyzed by DNA polymerase I, the ligation catalyzed by T4 DNA ligase and the digestion catalyzed by DNase I, along with comparisons of interfacial reaction efficiency when using AVS versus not using AVS. ( B ) Schematic diagram outlining the iEOS process on AVS. ( C ) Comparison of the stepwise yield, full-length yield, and deletion error rate with and without the use of AVS. ( D ) On the basis of NGS results, a synthetic step-by-step yield was determined through comparison with the target sequence (* P < 0.05; ** P ≤ 0.001; *** P ≤ 0.0001).

Article Snippet: Following this, a 10-μl polymerization reaction buffer was prepared by mixing Klenow fragment of DNA polymerase (1 U/μl M0210, NEB), 100 μM deoxycytidine triphosphate–Cy3, 5 mM deoxy-ribonucleoside triphosphate (dNTP) mix, 50 mM NaCl, 10 mM tris-HCl, 10 mM MgCl 2 , and 1 mM dithiothreitol (DTT) (pH 7.9).

Techniques: Ligation, Comparison, Sequencing