SAMPLE
The COLO829T melanoma cell line (ATCC CRL-1974, http://www.lgcstandards-atcc.org/products/all/CRL-1974) and COLO829BL peripherial blood cell line (ATCC CRL-1980, http://www.lgcstandards-atcc.org/products/all/CRL-1980) were obtained from ATCC and cultured as recommended.
METHODS
Library Preparation: Iso-Seq® Express Template Preparation for Sequel® and Sequel® II Systems (no part number yet)
Sequencing: Sequel System with Sequel Binding Kit 3.0 (101-500-400) and Sequel Sequencing Kit 3.0 (101-597-900)
Run time: 20 hrs pre-extension, 4 movie time per SMRT Cell
Analysis: SMRTlink 7.0 "IsoSeq" protocol, followed by mapping to hg38 reference genome and collapsed into non-redundant transcript set. Post-mapping filtering using SQANTI2 software (v2.7, https://github.com/Magdoll/SQANTI2)
Download Data: https://downloads-ap.pacbcloud.com/public/dataset/Melanoma2019_IsoSeq/
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