For the construction of snRNA-seq libraries, Chromium Single Cell 3′ Reagent Kits (10X Genomics; v.2 chemistry for human, chimpanzee, bonobo, gorilla, gibbon, macaque, marmoset, mouse and opossum; v.3 chemistry for platypus and chicken) were used according to the manufacturer’s instructions. Then 15,000 to 20,000 nuclei were loaded per lane in the Chromium microfluidic chips and complementary DNA was amplified in 12 PCR cycles. Sequencing was performed with NextSeq550 (Illumina) according to the manufacturer’s instructions using the NextSeq 500/550 High Output Kit v.2.5 (75 cycles) with paired-end sequencing (read lengths of read1 26 bp, read2 57 bp; index1 8 bp, roughly 170 to 380 million reads per library for v.2 chemistry; read lengths of read1 28 bp, read2 56 bp Index1 8 bp, roughly 247 to 306 million reads per library for v.3 chemistry) (Supplementary Table 2 ).
For bulk RNA-seq data generation, RNA was extracted using the RNeasy Micro kit (QIAGEN). The tissues were homogenized in RLT buffer supplemented with 40 mM DTT. The RNA-seq libraries were constructed using the TruSeq Stranded messenger RNA LT Sample Prep Kit (Illumina) as described in ref. 2 (link). Libraries were sequenced on Illumina NextSeq550 using a single-end run (read1 159 bp; index1 7 bp) with roughly 24–60 million reads per library (Supplementary Table2 ).
For bulk RNA-seq data generation, RNA was extracted using the RNeasy Micro kit (QIAGEN). The tissues were homogenized in RLT buffer supplemented with 40 mM DTT. The RNA-seq libraries were constructed using the TruSeq Stranded messenger RNA LT Sample Prep Kit (Illumina) as described in ref. 2 (link). Libraries were sequenced on Illumina NextSeq550 using a single-end run (read1 159 bp; index1 7 bp) with roughly 24–60 million reads per library (Supplementary Table
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