Additional file 12 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 12: Table S2. Primers for amplification of AL librar

Additional file 3 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 3: Figure S3. MCA maxima correlates with input titration: Amount of input DNA is correlated with the local maxima of the MCA determined derivative RFU of the input titrations of both Nextera (A) and AL (B) library prepared samples, with R2 equal to 0.807 and 0.842 respectivel

Additional file 1 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 1: Figure S1. Standard NGS and FA-NGS Workflow Comparison: Side-by-side comparison of the Standard NGS workflow (left) and modified FA-NGS workflow (right) highlights how FA-NGS can save time and hands on steps in preparing NGS librarie

Additional file 7 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 7: Figure S7. Comparison of percent reads between Nextera and AL libraries shows similarities in output from two distinct NGS workflows: The distributions of Nextera (blue) and AL (red) libraries of percent reads are overlaid to highlight the similarities (p-value = 1) of sequencing output from these methods. The range of percent reads for the Nextera library (blue) was 0.39–1.95, with a mean of 1.04 and a standard deviation of 0.43. The range of percent reads for the AL library (red) was 0.25–2.89, with a mean of 1.04 and a standard deviation of 0.

Additional file 11 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 11: Table S1. Sequencing analysis for Nextera library: alignment analysis was performed using embedded MiSeq Reporter softwar

Additional file 8 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 8: Figure S8. Percent difference from sequence pooling of Nextera and AL libraries: The frequency of percent differences from the expected percent reads per sample (1.04) is represented as a histogram for the Nextera library (A), AL library (B

Additional file 10 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 10: Figure S10. Continuous fluorescence measurements of qPCR: RFU values per cycle number are plotted for 96 plasmid libraries (A) and 96 gDNA libraries (B

Additional file 5 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 5: Figure S5. Melting curve analysis of Nextera prepared plasmids: The melting curve plot (temperature vs. negative derivative of fluorescence (−dF/dT)) of every well from the Nextera library is plotted. From left to right, the plasmids tested are pXMJ19, pskb3-CopR1598, pGEN-292, pms612

Additional file 6 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 6: Figure S6. Melting curve analysis of AL prepared gDNA: The melting curve plot (temperature vs. negative derivative of fluorescence (−dF/dT)) of every well from the AL-gDNA library is plotted. From quadrant 1–4, the input concentrations are 500 pg, 250 pg, 125 pg, 62.5 p

Additional file 9 of Fluorescent amplification for next generation sequencing (FA-NGS) library preparation

Additional file 9: Figure S9. Sequencing quality scores of Nextera and AL libraries: The percentage of bases with ≥ Q30 quality score for PhiX Control Library and for Nextera and AL libraries demonstrates sequencing quality for FA-NGS librarie