From: Targeted sequencing library preparation by genomic DNA circularization
Experiment | 1 | 2 | 3 | 4 | 5 |
---|---|---|---|---|---|
Hybridization temperature (°C) | 60 | 55 | 50 | 55 | 60 |
Number of PCR cycles | 25 | 25 | 25 | Direct | 25 |
Sequencing read length (mate pair versus single) | 42 | 42 | 42 | 42 | 42 by 42 |
Total reads | 12,542,683 | 15,605,713 | 12,435,664 | 1,232,093 | 33,750,858 |
Mapped reads a | 8,576,700 | 13,415,111 | 7,381,662 | 11,726 | 31,325,015 |
Captured on-target fragment-end reads c | 7,560,090 | 11,105,527 | 6,330,012 | 8,488 | 30,994,237 |
Captured off-target reads (as % of mapped) | 1,016,610 (12%) | 2,309,584 (17%) | 1,051,650 (14%) | 3,238 (28%) | 660,937 (2%) |
On-target fragment-end sequences (bases) c | 4,410 | 4,410 | 4,410 | 4,410 | 8,778 |
Captured on-target fragment-ends with 1× (bases) c, d (as % of on-target region-of-interest) | 3,145 (71%) | 3,340 (76%) | 3,044 (69%) | 2,809 (64%) | 6,628 (75%) |
Captured on-target fragment-ends with 30× (bases)b, c (as % of on-target region-of-interest) | 2,932 (66%) | 3,128 (71%) | 2,961 (67%) | 2,160 (49%) | 6,460 (73%) |
Average on-target fragment-end fold-coverage | 72,001 | 105,767 | 60,286 | 81 | 149,100 |