Table 2.
Customized Illumina Oligos for Small RNA v1.5 Libraries. Tm calculations for ave size 200bp (lib before index) and average %GC in 20 mM NaCl. Tm = 81.5 + 16.6 × log10(cation conc) + 0.41 × pctGC − (675.0 / length). Tm calculations for PCR reactions were done using the IDT OligoAnalyzer 3.1 with the following Phusion PCR salts; 50mM KCl, 2mM MgCl2.
| Custom smRNA v1.5 Oligos | Tm-DSN HYB | Tm-PCR | %GC |
|---|---|---|---|
| RS-TS-5’ adapter | 60.4 | 50.0 | |
| RS-TS-5’/PCR2D overlap | 61.9 | 50.0 | |
| RS-TS-5’ end (final) | 61.3 | 53.1 | |
| RS-TS-3’ adapter | 55.2 | 54.5 | |
| RS-TS-3’ PCR1A | 55.2 | 54.5 | |
| RS-TS-3’/PCR1A overlap | 72.3 | 54.5 | |
| RS-TS-3’ PCR-I-1 | 64.5 | 53.1 | |
| RS-TS-3’/PCR-I-1 overlap | 72.3 | 54.5 | |
| RS-TS-3’ end (final) | 64.5 | 53.1 | |
| Targets | Tm-DSN HYB | %GC | |
| 5.8S rRNA | 72.4 | 57.0 | |
| 5S rRNA | 76.4 | 59.5 | |
| 18S rRNA | 71.9 | 56.1 | |
| 28S rRNA | 77.2 | 69.4 | |