GSM6805893: Carotid artery, Day 0 (pre-injury), Replicate 4; Mus musculus; OTHER
Frozen tissue sections were mounted on special LCM membrane slides and stained immunohistochemically with Hematoxylin and Eosin to detect carotids and elastic laminas. Carotid sections containing outer elastic lamina and neointima were then cut and catapulted into the lid of a collection tube by the laser. After collection of up to 70 sections, total RNA was extracted using an RNeasy Micro Kit (Qiagen, Hilden, Germany). nanoMACEseq was performed for further analysis. The library preparation was conducted by GenXPro. 3'mRNA libraries were prepared using GenXPros “Rapid MACE-Seq Kit” for low-input mRNA according to the manual of the manufacturers. Briefly, RNA was fragmented to an average size of 350 bps, followed by poly-A specific cDNA synthesis, pooling and amplification by PCR using the minimum number of cycles as described in the manual. The PCR product was purified by SPRI purification. The final product of 200-400 bps was quality controlled on a Perkin Elmer LabChip GXII, the concentration was measured on Qbit.