Collection Tube Buffer
| Reagents | Volume per tube (µL) |
|---|---|
| dNTPs (10 mm) | 1.2 |
| First-strand buffer (5×; supplied with Superscript III [Invitrogen]) | 2.4 |
| DTT (100 mm; supplied with Superscript III) | 1.2 |
| Superase in RNase inhibitor (20 U/µL; Life Technologies) | 0.5 |
| dT-T7 oligo (100 ng/µL) | 0.3 |
All solutions, reagents, and plastic ware used to prepare this buffer should be RNase-free and designated for single-cell analysis only. The dT-T7 oligo sequence is GGAGGCCGGAGAATTGTAATACGACTCACTATAGGGAGACGCGTGTTTTTTTTTTTTTTTTTTTTTTTTV, where V is any nucleotide except T. Prepare this buffer in 1.5-mL microcentrifuge tubes. If prepared in advance, freeze at −20°C and add the RNase inhibitor immediately before use.
- © 2015 Cold Spring Harbor Laboratory Press
