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Table 2 Occurrence of chimeric BC–ROI combinations under different PCR conditions for high-diversity MPRA libraries

From: Optimized PCR conditions minimizing the formation of chimeric DNA molecules from MPRA plasmid libraries

Type of PCR

Elongation time, sa

MPRA library

Total read count

Reads with chimeric BC–ROI combinations, %

name

sizeb

R#1

R#2

R#1

R#2

emulsion

10

library-71

22,621

526,056

497,217

0.580

0.555

30

library-71

51,650

594,511

514,941

0.342

0.310

30

library-83

38,156

371,889

456,380

0.261

0.304

conventional

30

library-71

85,249

776,878

784,138

0.336

0.337

30

library-83

52,654

788,801

809,402

0.295

0.310

  1. aIndicated elongation time was used in both rounds of PCR
  2. bThe number of genuine BCs found in both replicates
  3. R#1 and R#2 stand for replicate 1 and replicate 2, respectively