Barcode_capture_library
To enrich the library for barcode sequences, we first performed PCR with biotinylated primers to generate a dsDNA biotinylated bait containing the 120bp of sequence immediately 3 of the barcode sequence in our expression vector. We isolated the bait DNA from the PCR using a 3X volume of ProNex beads, and eluted in 10 mM Tris pH 8.0 buffer. Next, we used 200 fmol of bait DNA as a custom probe in the Parse Gene Capture kit, following the manufacturers protocol aside from this substitution. After qPCR to verify the barcode enrichment, the resulting library was sequenced on one lane of a NovaSeq X Plus 10B PE150 flowcell.