![\"\u5355\u901a\u9053\u548c\u591a\u901a\u9053\u79fb\u6db2\u7ba1\"](\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-89.png\")
Swift2S\u2122 Turbo DNA Library Kit (left). All these experiments used the thermometer module required for active cooling of the enzyme and the magnetic module required for bead cleaning. Thermal cycler modules can be added to the OT-2 deck for full automation. The Swift2S\u2122 Turbo protocol includes enzymatic shearing of input DNA, adapter ligation, indexing, or optional PCR. Use SPRI cleanup after ligation and PCR steps.<\/p>\n\n\n\n
Results Along with the time savings of automation, Swift2S\u2122 prepared Turbo libraries on OT-2 are robust and comparable to manually generated libraries. To illustrate the unbiased performance of Swift2S\u2122 Turbo OT-2, the library input was 100 ng and a bacterial mixture of three strains with GC content (33 -71%) was used. The library was prepared by an experienced developer of both the Swift2S\u2122 Turbocharger and the Opentrons OT-2 reagent DE, used to slow down the fragmentation of enzymatic reactions in chambers for longer periods of time in automated liquid handling The process temperature (2) is added to the fragmentation master mix program. Thus, comparable to the library size, the yield was 85% compared to the manually prepared library on the OT-2 version (Figure 2). The CV of the library yield was 14% across 8 technical replicates prepared on OT-2.<\/p>\n\n\n\n
Sequencing metrics are also comparable when prepared manually and on the OT-2. For example, 9% of reads mapped to the reference genome in the bacterial mixture, indicating no contamination or reduced infidelity in libraries prepared on OT-2 (Figure 3). Comparable percentages of duplicates observed in manual and automated preparations indicate that DNA is retained throughout the SPRI cleanup operations performed on OT-2 (Figure 3). Comparable performance and sequence representation were further demonstrated by consistent coverage and relative abundance between automated and manual library preparers (Figure 4).<\/p>\n\n\n\n Figure 3. Manually compiled library of comparable ranking indicators. Manually prepared libraries are shown in yellow, libraries prepared on the opentronOT-2 are shown in blue. The left and right panels show similar read volumes and percentages of duplicates aligned to the reference genome. These metrics indicate that no contamination or loss of input DNA affects library complexity.<\/p>\n\n\n\n Figure 4. The relative abundance and coverage of the hand-prepared library and the library on the opentronOT-2 were comparable. Libraries were normalized to have the same number of sequencing reads (1.9 million reads) for comparative analysis. <\/em><\/p>\n\n\n\n Conclusion Automated library preparation reduces the potential for human error and reduces hands-on time and reagents without sacrificing the quality of library preparation. This automated Swift2S\u2122 Turbo protocol on the OT-2 platform enables robust preparation of 8 or more libraries in 2 to 3 hours with minimal yield variation between technical replicates. .Reagent DE (2) is used for automatic settings at room temperature to slow down fragmentation. .Libraries prepared using Opentrons OT-2 were comparable and reproducible (CV=14%). .Similar sequencing metrics were observed for manually prepared libraries and libraries prepared on OT-2, with comparable complexity, bias, and coverage.<\/p>\n","protected":false},"excerpt":{"rendered":" Summary The Swift2S\u2122 Turbo DNA Library Kit is automated […]<\/p>\n","protected":false},"author":5,"featured_media":3033,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"inline_featured_image":false,"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"categories":[],"tags":[],"class_list":["post-3032","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry"],"yoast_head":"\n![\"\u624b\u52a8\u548cOpenteronsOT-2\"](\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-90.png\")
![\"\u624b\u52a8\u7f16\u5236\u7684\u53ef\u6bd4\u8f83\u7684\u6392\u5e8f\u6307\u6807\u5e93\"](\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-91.png\")
<\/figure>\n\n\n\n![\"\u624b\u5de5\u5236\u5907\u7684\u5e93\u548copentronOT-2\u4e0a\u7684\u5e93\u7684\u76f8\u5bf9\u4e30\u5ea6\u548c\u8986\u76d6\u7387\u76f8\u5f53\"](\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-92.png\")
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