Combination of direct boiling and glass beads increases the purity and accuracy of bacterial DNA extraction

Abstract

Extraction of DNA is a key step in molecular biology experiments and important for counting tiny microbial individuals. Direct boiling and mechanical cell lysis like glass beads are two independent physical extraction methods, thus crossing the barriers of thresholds of magnitude in popular reagent kits or traditional spread plate method when non‐equilibrium phenomenon is ongoing. The two approaches above were combined to generate a new one. In three typical microbial species, direct boiling with glass beads significantly increased the purity of DNA solution compared with some other methods (p < 0.05). The qPCR results of them were closer to direct microscopy counting than some other methods. Therefore, it provides a new choice in extracting bacterial DNA for specific circumstances.

Simultaneous Detection of Helicobacter pylori and Clarithromycin Resistance Mutations Using RAA-CRISPR/Cas13a Assay

“…The simplicity, good specificity and sensitivity provide the possibility for large-scale screening and on-site testing. Given the advancements and progress in bacterial nucleic acid extraction technology, such as direct boiling, mechanical lysis, 25 , 26 the integration of SHIELD assay with streamlined and rapid nucleic acid extraction techniques needs further research.…”

Cure or Curse? Simulation Indicates that Microbes Proliferate under Disinfection Measures in the Space Station

“…DNA was subsequently extracted from the solution above (with PMA or not) by direct boiling with glass beads (Li et al, 2023). 0.1g 0.1mm glass beads (Cat.…”

“…The primers, their qPCR standard curves, their amplicons and the composition of qPCR reaction buffer were the same as our previous study (Li et al, 2023). 2×SYBR Green PCRmix (Cat.…”