A | B | |
NTC | No-template control | |
EPC | External positive control | |
Sample | wastewater sample |
COMPONENT | VOLUME (µL) | 10.4 | |
End Prep Reaction Mixture | 30 | - | |
Diluted or undiluted NEBNext UMI Adaptors for Illumina | 1.25 | - | |
(red) NEBNext Ultra II Ligation Master Mix | 15 | 156 | |
(red) NEBNext Ligation Enhancer | 0.5 | 5.2 | |
Total Volume | 46.75 |
A | B | C | D | E | |
Sequencer type | Diluent | Recommended Concentration | Minimum Concentration | Minimum Volume (uL) | |
MiSeq and NovaSeq | EB-Tween | >5nM* | >2.5nM | 50 (1 lane) |
A | B | C | D | E | F | G | H | I | J | |
Sample ID | Total library DNA amount (µg) | Remaining volume (µL) | Concentration (ng/µL) | Volume ratio to reach lowest concentration | Scaled volume to mix (µL)* | Rounded volume (µL) | DNA amount in mixed volume (ng) | Mixed DNA concentration (ng/µL) | Mixed DNA concentration (nM) | |
s001 | 1.9942 | 29.5 | 67.6 | 0.4452662722 | 6.2337278107 | 6.23 | 421.148 | |||
s002 | 2.22725 | 29.5 | 75.5 | 0.3986754967 | 5.5814569536 | 5.58 | 421.29 | |||
s003 | 2.03845 | 29.5 | 69.1 | 0.4356005789 | 6.0984081042 | 6.1 | 421.51 | |||
s004 | 2.65795 | 29.5 | 90.1 | 0.3340732519 | 4.6770255272 | 4.68 | 421.668 | |||
s005 | 2.1594 | 29.5 | 73.2 | 0.4112021858 | 5.7568306011 | 5.76 | 421.632 | |||
s006 | 3.363 | 29.5 | 114 | 0.2640350877 | 3.6964912281 | 3.7 | 421.8 | |||
s007 | 3.5105 | 29.5 | 119 | 0.2529411765 | 3.5411764706 | 3.54 | 421.26 | |||
s008 | 0.88795 | 29.5 | 30.1 | 1 | 14 | 14 | 421.4 | |||
s009 | 1.1859 | 29.5 | 40.2 | 0.7487562189 | 10.4825870647 | 10.48 | 421.296 | |||
s010 | 1.1918 | 29.5 | 40.4 | 0.745049505 | 10.4306930693 | 10.43 | 421.372 | |||
Total volume | 70.5 | 4214.376 | 59.7783829787 | 164.6787409882 |