title: GSM7509399: Left Cartilage 489 Exp Sept_2021 [489L_S9_L001]; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using the Monarch Total RNA Miniprep Kit (New England Biolabs), following manufacturer instructions. From this, RNA-Seq of messenger RNAs (mRNAs) was performed using a custom in-house multiplex method, which allowed us to sequence up to 24 different samples in the same sequencing lane. Briefly, samples were given a unique i7 index (together with UMI) during individual polyA-priming and first-strand synthesis, which also added a template-switch sequence to the 5' end. Samples were then pooled and amplified using P7 and an oligo which binds the template-switch sequence. Final library construction was completed by tagmentation and addition of P5 (with i5 index) by PCR.