Erratum: Application of RNA Interference in the Pinewood Nematode, Bursaphelenchus xylophilus

Resumen

An erratum was issued for: Application of RNA Interference in the Pinewood Nematode, Bursaphelenchus xylophilus. The Abstract and Protocol section was updated.

Resumen

This corrects the article 10.3791/63645-v

Protocolo

An erratum was issued for: Application of RNA Interference in the Pinewood Nematode, Bursaphelenchus xylophilus. The Abstract section was from:

This paper outlines a new protocol for RNAi of the ppm-1 gene in B. xylophilus, which has been reported to play crucial roles in the development and reproduction of other pathogenic nematodes. For RNAi, the T7 promoter was linked to the 5′-terminal of the target fragment by polymerase chain reaction (PCR), and double-stranded RNA (dsRNA) was synthesized by in vitro transcription. Subsequently, dsRNA delivery was accomplished by soaking the nematodes in a dsRNA solution mixed with synthetic neurostimulants. Synchronized juveniles of B. xylophilus (approximately 20,000 individuals) were washed and soaked in dsRNA (0.8 µg/mL) in the soaking buffer for 24 h in the dark at 25 °C.

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This paper outlines a new protocol for RNAi of the ppm-1 gene in B. xylophilus, which has been reported to play crucial roles in the development and reproduction of other pathogenic nematodes. For RNAi, the T7 promoter was linked to the 5′-terminal of the target fragment by polymerase chain reaction (PCR), and double-stranded RNA (dsRNA) was synthesized by in vitro transcription. Subsequently, dsRNA delivery was accomplished by soaking the nematodes in a dsRNA solution mixed with synthetic neurostimulants. Synchronized juveniles of B. xylophilus (approximately 20,000 individuals) were washed and soaked in dsRNA (0.8 µg/μL) in the soaking buffer for 24 h in the dark at 25 °C.

The Protocol section was from

3. RNAi by soaking

1. Mix 4 µL of 5x soaking buffer (0.05% gelatin, 5.5 mM KH₂PO₄, 2.1 mM NaCl, 4.7 mM NH₄Cl, 3 mM spermidine) with the dsRNA and ddH₂O to produce a total volume of 20 µL and a final RNA concentration of 0.8 µg/mL.

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3. RNAi by soaking

1. Mix 4 µL of 5x soaking buffer (0.05% gelatin, 5.5 mM KH₂PO₄, 2.1 mM NaCl, 4.7 mM NH₄Cl, 3 mM spermidine) with the dsRNA and ddH₂O to produce a total volume of 20 µL and a final RNA concentration of 0.8 µg/μL.