BioStringTM Total RNA Plus Kit was evaluated as a rapid silica column-based total RNA extraction workflow with integrated genomic DNA removal. The kit uses a DNA Mini Column before RNA binding, enabling total RNA purification from cultured cells and animal tissues without phenol / chloroform extraction or isopropanol precipitation. The purified RNA is intended for downstream applications such as RT-PCR, Northern blotting, poly(A)+ RNA purification, nuclease protection and in vitro translation.
In protein- and lipid-rich tissue testing, RNA was extracted from 10 mg chicken liver and 10 mg chicken brain samples. From 10 mg chicken liver input, the kit produced RNA yields of 45.46–48.24 µg, with A260/280 values of 2.11–2.12 and A260/230 values of 1.75–2.23. From 10 mg chicken brain input, RNA yields were 4.29–4.55 µg, with A260/280 values of 2.13–2.18 and A260/230 values of 1.90–2.15 under the tested conditions.
A comparison with a reference column-based RNA extraction workflow was performed using chicken liver input amounts of 5 mg, 10 mg, 20 mg and 40 mg. With 40 mg chicken liver input, R4111 produced RNA yields of 147–156 µg, compared with 147 µg from the reference workflow. With 20 mg input, R4111 produced 95–98 µg RNA, compared with 84 µg from the reference workflow. With 10 mg input, R4111 produced 41–47 µg RNA, compared with 37 µg from the reference workflow, while 5 mg input produced 19–21 µg RNA under the tested conditions.
Across the tested chicken liver input range, maintained A260/280 values of approximately 2.11–2.15 and A260/230 values of approximately 2.07–2.28. Agarose gel electrophoresis showed intact RNA band patterns and no obvious genomic DNA contamination, supporting the effectiveness of the gDNA removal column for routine DNA reduction before RNA purification.
The kit was also evaluated in a rapid cultured-cell extraction workflow using 5 × 106 cultured cells per sample. Six cell samples were processed in approximately 30 minutes, and electrophoresis analysis showed clear total RNA bands with good integrity. These results support R4111 as a fast total RNA extraction workflow for routine animal tissue and cultured-cell samples where reduced genomic DNA background and short hands-on processing time are required