Using single-worm RNA sequencing to study C. elegans responses to pathogen infection

BMC Genomics

Table 1 Three conditions tested for the mechanical lysis of worms

Condition	Processing before shearing	Device of shearing
1	No processing	TissueLyser LT^b
2	Incubated in Lysis Buffer^a at 65 °C for 10 min, 85 °C for 1 min, then held at 4 °C in a thermocycler	TissueLyser LT^b
3	Incubated in Lysis Buffer^a at 65 °C for 10 min, 85 °C for 1 min, then held at 4 °C in a thermocycler	QIAshredder spin column^c

^aLysis Buffer was composed of 50 mM KCl, 10 mM Tris–HCl, pH 8.3, 2.5 mM MgCl₂, 0.45% Triton X-100, 0.45% Tween-20, 0.11% gelatin (w/v), 100 µg/mL proteinase K, and 200 units of RNasin Ribonuclease Inhibitor
^bA TissueLyser LT was used following the “Purification of RNA or Multiple Analytes form Animal and Human Tissues” protocol in the manufacturer’s manual
^cThe QIAshredder spin column was centrifuged at 16,873 × g for 2 min

ISSN: 1471-2164