Materials
| Name | Company | Catalog Number | Comments |
| ROSALUC Mice | made in house | frozen sperm available upon request | |
| R26-iPSC mice | made in house | frozen sperm available upon request | |
| Vessel probe | Fine Science Tools | 10160-13 | to check for copulation plugs |
| M2 medium | Sigma-Aldrich | M7167 | make aliquots and store at -20 °C |
| Fine forceps (Dumont #5 Standard tip Student forceps) | Fine Science Tools | 11251-10 | spray with 70% EtOH before use (do not autoclave) |
| 23 G needles | Fine-ject | 8697 | |
| 1-mL syringes | Soft-ject | 6680 | |
| 60-mm bacterial grade plates (for flushing) | Gosselin | BB60-01 | |
| Mouth pipette | made in house | see discussion | |
| Mouse embryonic fibroblasts (MEFs, TgN (DR4)1 Jae strain) | ATTC | SCRC-1045 | |
| TgN (DR4)1 Jae mice | The Jackson Laboratory | 3208 | |
| Mitomycin C | Sigma-Aldrich | M0503 | |
| Phosphate buffered saline (PBS) without calcium or magnesium | Gibco | 14190-094 | |
| Tg(DR4)1Jae/J mice | JAX | 3208 | mice that contain four drug-selectable genes and DR4 MEFS confers resistance to neomycin, puromycin, hygromycin and 6-thioguanine |
| 0.1% Gelatin | Sigma-Aldrich | G1393 | Dissolve 0.5 g in 500 mL distilled water, autoclave and store at 4 °C. |
| Trypsin (0.25%) | Gibco | 25200-056 | |
| 2 μM pluripotin | Cayman Chemical | 10009557 | |
| pRMCE-DV1 | BCCM/LMBP collection | LMBP 08870 | public available from the BCCM/LMBP collection (http://bccm.belspo.be) |
| cre-excised pRMCE-DV1 | BCCM/LMBP collection | LMBP 08195 | public available from the BCCM/LMBP collection (http://bccm.belspo.be) |
| pCAG-FlpE-IRES-Puro-pA | Addgene | 20733 | |
| heat-shock competent DH5α bacteria | made in house | ||
| Gateway pDONR221 vector | Thermo Fisher | 12536-017 | contains kanamycin-resistance gene |
| BP clonase II mix | Thermo Fisher | 11789-020 | |
| LR clonase II mix | Thermo Fisher | 11791-020 | |
| Luria Broth (LB) | |||
| Ampicillin | |||
| Applied Biosystems 3730XL DNA Analyzer | Thermo Fisher | 3730XL | |
| G418 | Thermo Fisher | 11811-023 | |
| Lipofectamine 2000 transfection reagent | Thermo Fisher | 11668027 | |
| Lipofectamine LTX transfection reagent | Thermo Fisher | 15338100 | |
| Effectene transfection reagent | Qiagen | 301425 | |
| GATEWAY pENTR 1A vector | Thermo Fisher | A10462 | recombination-compatible vector |
| mouse monoclonal anti-p120ctn antibody | BD Transduction Laboratories | 610134 | |
| mouse monoclonal anti-Ecadherin antibody | BD Transduction Laboratories | 610181 | |
| General equipment | |||
| Sterile dissection tools | fine scissors and forceps for dissecting the uterus | ||
| Sterile pipettes: 5 mL, 10 mL and 25 mL | |||
| 15-mL and 50-mL conical centrifuge tubes | |||
| 96-well culture plates V-shaped bottom and flat bottom) | |||
| Culture dishes: 24 wells, 12 wells and 6 wells | |||
| Multichannel pipettes (to pipette 30, 50, 100 and 200 μL) | |||
| Sterile multichannel reservoirs | |||
| Access to a laminar air flow | |||
| Access to an incubator at 37 °C with 5% CO2 | |||
| Access to an inverted microscope | |||
| Access to a bench-top centrifuge | |||
| Access to a stereo microscope with transmitted-light | |||
| Culture media | |||
| MEF Medium | stored at 4 °C; warm 30 min at 37 °C before use | ||
| Dulbecco’s modified Eagle’s medium (DMEM) | Gibco | 41965-062 | |
| 10% fetal bovine serum (FBS) | Sigma-Aldrich | F-7524 | |
| L-glutamine (2 mM) | Gibco | 25030-024 | |
| Sodium pyruvate (0.4 mM) | Gibco | 11360-039 | |
| penicillin (100 U/mL) | Gibco | 15140-122 | |
| streptomycin (100 µg/mL) | Gibco | 15140-122 | |
| SR-based mESC medium | stored at 4 °C; warm 30 min at 37 °C before use | ||
| DMEM/F12 | Gibco | 31330-038 | mixed in a 1:1 ratio |
| 15% knock-out serum replacement (SR) | Gibco | 10828–028 | |
| L-glutamine (2 mM) | Gibco | 25030-024 | |
| 0.1 mM non-essential amino acids | Gibco | 11140-050 | |
| penicillin (100 U/mL) | Gibco | 15140-122 | |
| streptomycin (100 µg/mL) | Gibco | 15140-122 | |
| β-mercaptoethanol (0.1 mM) | Sigma-Aldrich | M 3148 | |
| 2,000 U/mL recombinant mouse LIF | (IRC/VIB Protein Service facility) | ||
| FBS-based mESC medium (similar to SR-based mESC medium) | stored at 4°C; warm 30 min at 37°C before use | ||
| Knockout DMEM | Gibco | 10829-018 | |
| 15% FBS | Hyclone | SH30070.03E | |
| Differention Medium | stored at 4 °C; warm 30 min at 37 °C before use | ||
| Iscove's Modified Dulbecco's Medium (IMDM) | Gibco | 21980-032 | |
| 15% FBS | Hyclone | SH30070.03E | |
| 5% CD Hybridoma Medium(1x) liquid | Gibco | 11279-023 | |
| 2 mM L-glutamine | Gibco | 25030-024 | |
| 0.4 mM 1-thioglycerol | Sigma-Aldrich | M-6145 | |
| 50 μg/mL ascorbic acid | Sigma-Aldrich | A-4544 | |
| penicillin (100 U/mL) | Gibco | 15140-122 | |
| streptomycin (100 µg/mL) | Gibco | 15140-122 | |
| 2i | |||
| 1 μM Erk inhibitor PD0325901 | Axon Medchem | Axon 1408 | |
| 3 μM Gsk3 inhibitor CHIR99021 | Axon Medchem | Axon 1386 |
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