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- step-library-prep type Thing assertion.
- step-library-prep comment "Excludes RNase-cleaved transcripts (5'-OH and 3'-PO4 incompatible with ligation)" assertion.
- step-library-prep description "Selective sequencing of intact (non-RNase-cleaved) transcripts" assertion.
- step-library-prep substep "First-strand cDNA synthesis (M-MLV reverse transcriptase)" assertion.
- step-library-prep substep "3'-adapter ligation to hydroxyl ends" assertion.
- step-library-prep substep "5'-adapter ligation to phosphorylated ends" assertion.
- step-library-prep substep "Cap removal (RNA 5'-pyrophosphohydrolase)" assertion.
- step-library-prep substep "PCR amplification (15 cycles, high-fidelity polymerase)" assertion.
- step-library-prep substep "Size selection: 10-220 nt on agarose gel" assertion.
- step-library-prep name "Custom Library Preparation" assertion.