Table 2.
Reagents details.
| |
Antibodies used for immunocytochemistry/flow-cytometry |
|||
|---|---|---|---|---|
| Antibody | Dilution | Company Cat # | RRID | |
|
| ||||
| Pluripotency Markers | Mouse anti-OCT3/4 | 1:30 | Santa Cruz Cat# sc-5279 | RRID: AB_628051 |
| Rabbit anti-NANOG | 1:100 | Proteintech Cat# 14295-1-AP | RRID: AB_1607719 | |
| Secondary Antibodies | Alexa Fluor Plus 647 Goat anti-Mouse IgG | 1:1000 | Invitrogen Cat# A32728 | RRID: AB_2633277 |
| Alexa Fluor Plus 647 Goat anti-Rabbit IgG | 1:1000 | Invitrogen Cat# A32733 | RRID: AB_2633282 | |
| Primers | ||||
|
| ||||
| Target | Size of band | Forward/Reverse primer (5′-3′) | ||
|
| ||||
| Sendai Virus (PCR) | Sendai Virus Genome | 181 bp | GGATCACTAGGTGATATCGAGC/ACCAGACAAGAGTTTAAGAGATATGTATC | |
| Transgene (PCR) | c-Myc | 532 bp | TAACTGACTAGCAGGCTTGTCG/TCCACATACAGTCCTGGATGATGATG | |
| House-Keeping Gene (PCR) | GAPDH | 238 bp | GAGTCAACGGATTTGGTCGT/TTGATTTTGGAGGGATCTCG | |
| Pluripotency Markers (qPCR) |
NANOG
SOX2 |
237 bp 155 bp |
AAGGTCCCGGTCAAGAAACAG/CTTCTGCGTCACACCATTGC GCCGAGTGGAAACTTTTGTCG/GGCAGCGTGTACTTATCCTTCT |
|
| Ectoderm Markers (qPCR) | PAX6 | 111 bp | TGGGCAGGTATTACGAGACTG/ACTCCCGCTTATACTGGGCTA | |
| OTX2 | 179 bp | CAAAGTGAGACCTGCCAAAAAGA/TGGACAAGGGATCTGACAGTG | ||
| Endoderm Markers (qPCR) | SOX17 | 94 bp | GTGGACCGCACGGAATTTG/GGAGATTCACACCGGAGTCA | |
| FOXA2 | 83 bp | GGAGCAGCTACTATGCAGAGC/CGTGTTCATGCCGTTCATCC | ||
| Mesoderm Markers (qPCR) | TBX6 | 139 bp | CATCCACGAGAATTGTACCCG/AGCAATCCAGTTTAGGGGTGT | |
| MYH6 | 187 bp | CAAGTTGGAAGACGAGTGCT/ATGGGCCTCTTGTAGAGCTT | ||
| House-Keeping Gene (qPCR) | HPRT | 94 bp | TGACACTGGCAAAACAATGCA/GGTCCTTTTCACCAGCAAGCT | |
| DSP Amplification for Sequencing (PCR) | DSP | 1406 bp | GGGGCTCCAGGACTCCATCAG/GTCCTTCAGGATACACTCATCCCCC | |
| DSP Sequencing (Sanger) | DSP | GCGTCAGGTGCAGAACTTGG | ||