Reagent | Volume per Reaction | |
5x RT Buffer | 2μL | |
10mM dNTP | 1μL | |
Molecular Biology-Grade H₂O | 1μL | |
0.1M DTT | 0.5μL | |
SSIV RT | 0.5μL | |
TOTAL | 5μL |
Reagent | Volume per Reaction | |
5x Sequenase | 1μL | |
Molecular Biology-Grade H₂O | 3.8μL | |
Sequenase | 0.15μL | |
TOTAL | 4.95μL |
Reagent | Volume per Reaction | |
Sequenase dilution buffer | 0.45μL | |
Sequenase | 0.15μL | |
TOTAL | 0.6μL |
Reagent | Volume per Reaction | |
2X LongAmp Taq Master Mix | 44μL | |
SOL Primer B | 1μL | |
TOTAL | 45μL |
Step | Temperature | Time | Number of Cycles | |
Initialization | 98°C | 30 seconds | 1 | |
Denaturation | 94°C | 15 seconds | 30 | |
Annealing | 50°C | 20 seconds | ||
Extension | 68°C | 2 minutes | ||
Final Extension | 68°C | 10 minutes | 1 | |
Hold | 4°C | ∞ | -- |
Step | Temperature | Time | Number of Cycles | |
Hold | 4°C | -- | 1 | |
Incubation 1 | 37°C | 30 minutes | 1 | |
Incubation 2 | 65°C | 30 minutes | 1 | |
Hold | 4°C | ∞ | -- |
Reagent | Volume per Reaction | |
Molecular Biology-Grade H₂O | 15μL | |
Frag/AT Buffer | 4μL | |
Frag/AT Enzymes | 6μL | |
TOTAL | 25μL |
Step | Temperature | Time | Number of Cycles | |
Initialization | 98°C | 45 seconds | 1 | |
Denaturation | 98°C | 15 seconds | 12 | |
Annealing | 60°C | 30 seconds | ||
Extension | 72°C | 30 seconds | ||
Final Extension | 72°C | 1 minute | 1 | |
Hold | 4°C | ∞ | -- |
Number of Indexed Samples per Pool | Amount of Each Indexed Library per Pool | Total Mass per Pool | |
1 | 500 ng | 500 ng | |
2 | 500 ng | 1,000 ng | |
3 | 500 ng | 1,500 ng | |
4 | 375 ng | 1,500 ng | |
8 | 187.5 ng | 1,500 ng |
Reagent | Volume per Pool | |
Hybridization Mix | 20μL | |
Twist Respiratory Panel | 4μL | |
Molecular Biology-Grade H₂O | 4μL | |
TOTAL | 28μL |
Reagent | Volume per Pool | |
Dried, Indexed Library Pool | --- | |
Blocker Solution | 5μL | |
Universal Blockers | 7μL | |
TOTAL | 12μL |
A | B | C | D | |
Initialization | 98°C | 45 seconds | 1 | |
Denaturation | 98°C | 15 seconds | 15 | |
Annealing | 60°C | 30 seconds | ||
Extension | 72°C | 30 seconds | ||
Final Extension | 72°C | 1 minute | 1 | |
Hold | 4°C | ∞ | -- |
Reagent | Volume per Reaction | |
Streptavidin Binding Bead Slurry | 22.5μL | |
Amplification Primers, ILMN | 2.5μL | |
Equinox Library Amp Mix (2x) | 25μL | |
TOTAL | 50μL |