Genomic dna screentape kit

RNA extraction was done using the RNeasy Mini Kit (Qiagen, Hilden, Germany). After lysis of the cryosections in RLT buffer [+Dithiothreitol (DTT)] by vortexing, the kit was used according to the manual. The RIN^e-value is based on a mathematical algorithm based on the quantitative measurement of rRNA degradation. It has a high correlation to the RIN-values that can be measured by the Agilent Bioanalyzer instead of the TapeStation²⁵ . RIN^e was measured with the Genomic RNA-ScreenTape Kit, together with the RNA ScreenTape Sample Buffer- and Ladder on the Agilent TapeStation 4200, with the corresponding Analysis-Software, Version A.02.02 (all Agilent Technologies, Santa Clara, USA).
Extraction of DNA was done out of the cryo-sections with the QIamp DNA Micro Kit (Qiagen, Hilden, Germany), as described in the kits manual. DNA was eluted with 40 µl of RNAse free water. The DIN-value was measured with the Genomic DNA ScreenTape Kit on the same Agilent TapeStation 4200 (Agilent Technologies, Santa Clara, USA).
Sectioning and extracting was done on the same day for all 32 samples. RIN^e- and DIN-values were measured on the following day, with storing the extracted RNA and DNA at − 80 °C.

DNA was isolated from three separate cell incubations for all groups. PureLink Genomic DNA Mini Kit (Thermo Fisher, Waltham, MA, USA) was used for DNA isolation following the manufacturer’s instructions. The concentration and quality of the isolated DNA were measured using TapeStation 4510 (Agilent Technologies, Santa Clara, CA, USA) and the Genomic DNA ScreenTape kit (Agilent Technologies, Santa Clara, CA, USA). All samples showed DINs ≥ 9, which confirms the high quality of the genetic material. The bisulfate conversion was performed using the EZ DNA Methylation-Gold Kit (Zymo Research, Irvine, CA, USA) according to the manufacturer’s instructions. In total, 500 ng of DNA from each sample was used for conversion.