For RT² Profiler™ PCR Array 10 000 PKH26+ and 10 000 PKH26− cells (selected 12 days after PKH26 staining) were sorted into 100 μl Buffer RL (Norgen) as described before [13] . RNA was isolated with Single Cell RNA Purification Kit (Norgen) with On-Column DNA Removal (Norgen) followed by reverse transcription PCR using NCode Vilo (Invitrogen). RT² Profiler™ PCR Array (Qiagen) detecting murine CSC-related genes was performed using the vendor's protocol and StepOne Plus PCR (Applied Biosystems) cycler. Genes included in the array are listed in Supplementary Table 2.
On column dna removal
Manufactured by Norgen Biotek
Sourced in Canada
The On-Column DNA Removal is a laboratory equipment product designed to remove DNA from RNA samples. It functions by selectively binding and retaining DNA molecules while allowing RNA to pass through, facilitating the purification of RNA from DNA-containing samples.
Automatically generated - may contain errors
Lab products found in correlation
Ncode vilo, by Thermo Fisher Scientific (2 mentions)
Buffer rl, by Norgen Biotek (2 mentions)
Single cell rna purification kit, by Norgen Biotek (2 mentions)
Rt profiler pcr array, by Qiagen (1 mentions)
Steponeplus, by Thermo Fisher Scientific (1 mentions)
Rt2 profiler pcr array, by Qiagen (1 mentions)
2 protocols using on column dna removal
1
Profiling Murine Cancer Stem Cells
2
Quantifying Murine Cancer Stem Cells
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For the RT2 Profiler PCR Array analysis, we sorted 2000–10,000 MIC+ and MIC− B16-F10 cells in 100 μL of Buffer RL (Norgen, Thorold, Canada). Next, RNA was isolated using the Single Cell RNA Purification Kit (Norgen, Thorold, Canada) with the On-Column DNA Removal (Norgen, Thorold, ON, Canada) step according to the vendor’s protocol. RNA was eluted using 10 μL of DNase RNase-free H2O (this step was repeated 4 times to increase the RNA yield). Isolated RNA was entirely used for the RT-PCR using NCode Vilo (Invitrogen, Waltham, MA, USA), and obtained cDNA was diluted 6 times. The RT2 Profiler PCR Array (Qiagen, Germantown, MD, USA) detecting murine CSC-related genes was performed according to the vendor’s protocol using the Applied Biosystems StepOne Plus (Waltham, MA, USA) device. The list of genes is included in Table S1 .
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