CRISPR-Cas Systems Detection by PCR

DNA was extracted using the boiling method. The collected clinical isolates were tested for the presence of CRISPR-Cas systems (including type I-E^* and type I-E CRISPR systems) by polymerase chain reaction (PCR) using primers from a previous study (Li et al., 2018 (link)). PCRs were prepared using 2 × Hieff™ PCR Master Mix (with dye) (Yeasen) according to the manufacturer’s instructions (12.5 μL of Master Mix, 10 μM of each primer, and 1 μL DNA template, with a total volume of 25 μL).

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Hu Y., Jiang J., Wang D., Guo Q, & Wang M. (2023). Coexistence of blaKPC-IncFII plasmids and type I-E* CRISPR-Cas systems in ST15 Klebsiella pneumoniae. Frontiers in Microbiology, 14, 1125531.

Publication 2023

2 × Hieff™ PCR Master Mix (with dye)

Crispr Crispr cas systems Polymerase chain reaction Primer

Corresponding Organization : Fudan University

Other organizations : Gansu Provincial Hospital

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Variable analysis

independent variables

Presence of CRISPR-Cas systems (including type I-E* and type I-E CRISPR systems)

dependent variables

Outcomes of the polymerase chain reaction (PCR) testing for the presence of CRISPR-Cas systems

control variables

Boiling method used for DNA extraction
Primer sequences from the previous study (Li et al., 2018)
Concentration of Master Mix (2x Hieff™ PCR Master Mix)
Concentration of primers (10 μM)
Volume of DNA template (1 μL)
Total reaction volume (25 μL)

controls

No positive or negative controls were explicitly mentioned in the information provided.

Annotations

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