Table 3.
SmartSeq2, library preparation
| Type | Ingredient | Ratio | Manufacturer | Identifier |
|---|---|---|---|---|
| Lysis buffer | H2O | 1:53 | NA | |
| Recombinant RNase inhibitor | 1:40 | Takara | 2313A | |
| ERCC 1:600,000 | 1:40 | In-house | ||
| 10% Triton X-100 | 1:50 | Sigma | 93443 | |
| 10 mm dNTP | 1:4 | In-house | ||
| 100 µm dT | 1:40 | In-house | ||
| Reverse transcription mix | SMARTScribe | 1:6.32 | Takara | 639538 |
| Recombinant RNase inhibitor | 1:24 | Takara | 2313A | |
| 5× first strand buffer | 1:3 | Takara | 639538 | |
| 100 mm DTT | 1:12 | Promega | P117A | |
| 5 m betaine | 1:3 | Sigma | B0300 | |
| 1 m MgCl2 | 1:100 | Sigma | M1028 | |
| 100 µm TSO | 1:60 | In-house | ||
| H2O | 1:43 | NA | ||
| Preamplification mix | H2O | 1:7 | NA | |
| Kapa HiFi HotStart ReadyMix (2×) | 1:1.2 | Kapa Biosystems | KK2601 | |
| 10 µm ISPCR primer | 1:60 | In-house | ||
| λ Exonuclease | 1:133 | New England Biolabs | M0262S | |
| Tn5 mix | H2O | 1:1.7 | NA | |
| TAPS-PEG | 1:3.6 | In-house | ||
| Tn5 stock | 1.7.2 | In-house | ||
| PCR/bar code mix | H2O | 1:1.47 | NA | |
| 5× buffer | 1:3.9 | KAPA | KB2500 | |
| dNTPs | 1:26 | KAPA | KN1009 | |
| Hifi | 1:39 | KAPA | KE2004 | |
NA: Not Applicable.