PMC

Comparison of different homogenization strategies in young seeds and buds. (A) Young seeds without TRIzol addition, (B) buds without TRIzol addition, (C) young seeds with addition of 100 μl TRIzol before homogenization, (D) buds with addition of 100 μl TRIzol before homogenization, (E) Impact of homogenization strategy on RNA integrity. Marker—Invitrogen gene ruler 1 kb + (Cat. No. SM0311); left—undegraded RNA (low yield) isolated without TRIzol addition before, 1 min of homogenization of frozen plant tissue; middle—undegraded RNA (high yield) isolated with addition of 100 μl TRIzol before, 1 min of homogenization of thawed plant tissue; right—degraded RNA (high yield) isolated without TRIzol addition before, 3 × 1 min of homogenization in order to reach comparable tissue disruption efficiency in comparison with + TRIzol samples.

RNA integrity checking. Gel image generated by the automated electrophoresis system (Experion, Biorad). Lanes M – RNA molecular weight markers supplied with the Experion RNA StdSens analysis kit. Lanes PGTX beads – 1 μl of extracted RNA from distinct extractions using the PGTX solution combined with mechanical cell disruption with glass beads. Lanes Trizol beads – 1 μl of extracted RNA from distinct extractions using Trizol combined with mechanical cell disruption with glass beads. Lanes PGTX 95 – 1 μl of extracted RNA from distinct extractions using the PGTX solution combined with high temperature cell disruption. Lanes Trizol 95 – 1 μl of extracted RNA from distinct extractions using Trizol combined with high temperature cell disruption. Lanes Trizol std – 1 μl of extracted RNA from distinct extractions using Trizol according to general manufacturer's instructions. Lanes BPC – 1 μl of extracted RNA from distinct extractions using the BPC method. RNA integrity was kept for all extraction methods.

Real-time reverse transcriptase PCR (RT-PCR) results for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causing coronavirus disease 2019 (COVID-19). (A) FAM (N gene) quantitation. (a) RNA extraction and transport medium used TRIzol. (b) QIAamp kit for RNA extraction. (c) Viral transport medium with TRIzol RNA extraction. (d) Positive control. (B) VIC (ORF1ab) quantitation. (a) RNA extraction and medium transport were performed with TRIzol. (b) QIAamp RNA extraction with viral transport medium. (c) TRIzol extraction with viral transport medium. (d) Positive control. (C) FAM and VIC quantitation. (a) RNA extraction and medium transport were performed with TRIzol. (b) QIAamp RNA extraction with viral transport medium. (c) TRIzol extraction with viral transport medium. (d) Positive control. Three RNA extractions were performed for each patient.

Flow chart for protein extraction from Trizol. The protocol followed for protein extraction from the archived Trizol fractions is depicted as a flow chart. The extracted proteins were stored as aliquots at −80°C until further analysis.