title: GSM2773580: Icm02-W-s34; Homo sapiens; Bisulfite-Seq
library_construction_protocol: single cell or small amounts of cells were seeded into the lysis buffer using the mouth pipette, and DNAs were released after proteinase treatment at 50C and then bisulfite converted. After purification, DNAs were complemented with the biotinated random primer (5’-biotin-CTACACGACGCTCTTCCGATCTNNNNNNNNN-3’) and 50 units of klenow polymerase (3’ to 5’exo-, New England Biolabs). This random priming was repeated 5 times in total. The second strands were synthesized using another random primer (5’-AGACGTGTGCTCTTCCGATCTNNNNNNNNN-3’) and the final libraries were generated after 8-12 cycles PCR amplification with the Illumina universal PCR primer and Illumina indexed primer. For RNA-Seq, transcriptome of single cells or low amount of cells were amplified using a modified Smart-seq2. We reversely transcribed RNA molecules using a oligo (dT) primer anchored with 8bp unique molecular identifiers (UMI). Amplified cDNAs were fragmented into around 300 bp using a Covaris S2 and the sequencing libraries were constructed using a Kapa Hyper Prep Kit (Kapa Biosystems).
attributes.tag: GEO Accession
attributes.value: GSM2773580