Extended Data Fig. 2: Construction of RT&Tag libraries.
From: Profiling RNA at chromatin targets in situ by antibody-targeted tagmentation
Schematic showing how RT&Tag libraries are generated. During reverse transcription the oligo(dT)-ME-B fusion oligonucleotide binds to the poly(A) tail of RNA. Anchored oligo(dT) is used to ensure binding at the start of the poly(A) tail. Through the process of reverse transcription, the ME-B sequence gets appended to the cDNA. The RNA/cDNA hybrid then gets tagmented with ME-A loaded Tn5. Sequencing libraries are then amplified using primers complementary to the i5 and i7 sequences. The libraries are sequenced using 50 base pair single-end sequencing with the read originating from the i5 side.
