Reagents used in this study.
Source | Product ID | |
Cell lines | ||
C57BL/6J × CAST mESCs | From: Sun et al (2012) | N/A |
CAST × C57BL/6J mESCs | From: Sun et al (2012) | N/A |
129 × C57BL/6 mESCs | From: Mohn et al (2008) | N/A |
Chemicals, peptides, and recombinant proteins | ||
DMEM | Gibco | 21969-035 |
Nonessential amino acids | Gibco | 11140035 |
100 mM Sodium pyruvate | Gibco | 11360070 |
200 mM l-glutamine | Gibco | 25030024 |
FBS | Gibco | 10270106 |
β-Mercaptoethanol | Sigma-Aldrich | M-7522 |
Gelatin | Sigma-Aldrich | G-1890 |
Trypsin–EDTA | Gibco | 25300-054 |
Dulbecco’s PBS | Gibco | 14190 |
Trypsin (TPCK-treated) | Sigma-Aldrich | T8802 |
Retinoic acid | Sigma-Aldrich | R-2625 |
GSK3 inhibitor CHIR99021 | Calbiochem | 361559 |
MEK inhibitor PD 0325901 | Tocris | 4192 |
NDiff227 medium | Stem Cells | SCS-SF-NB-02 |
OptiMEM | Gibco | 31985070 |
Lipofectamine 3000 Transfection kit | Invitrogen | L3000015 |
cOmplete Protease Inhibitor Cocktail | Roche | 11836145001 |
Proteinase K | Roche | 3115879001 |
SuperScript III | Life Technologies | 18080085 |
Anti-FLAG M2 magnetic beads | Sigma-Aldrich | M8823 |
3× FLAG peptide | Sigma-Aldrich | F4799-25MG |
Dynabead M-280 Streptavidin | Invitrogen | 11206D |
RNace-It Ribonuclease Cocktail | Agilent | 400720 |
TSAP Thermosensitive Alkaline Phosphatase | Promega | M9910 |
RNasin Ribonuclease Inhibitor | Promega | N2115 |
Recombinant RNasin Ribonuclease Inhibitor | Promega | N2511 |
miR-cat 33 conversion oligo pack | IDT | Custom order |
T4 RNA Ligase 1 (ssRNA Ligase) | NEB | M0204L |
T4 PNK, T4 polynucleotide kinase | NEB | M0201L |
Hybond-C Extra membrane | GE Healthcare | RPN303E |
Kodak BioMax MS autoradiography film | Kodak | 8222648 |
MetaPhor agarose | Lonza | 50180 |
NuPAGE 4–12% (wt/vol) polyacrylamide Bis-Tris gels | Life Technologies | NP0335 |
NuPAGE LDS sample buffer 4× | Life Technologies | NP0007 |
NuPAGE SDS-MOPS running buffer | Life Technologies | NP0001 |
NuPage transfer buffer | Life Technologies | NP00061 |
MinElute Gel Extraction kit | Qiagen | 28604 |
Proteinase K | Roche | 03115836001 |
RNase H | NEB | M0297L |
TaKaRa long and accurate (LA) Taq | Clontech | RR002M |
γ32P-ATP 0.5 mCi 18.5 MBq Spec act. >6,000 Ci/mmol | Hartman | SRP-501 |
RCC RNA Spike-In Mix 1 | Ambion | 4456740 |
Single Cell RNA Purification kit | Norgen | 51800 |
Recombinant DNA | ||
SpCas9-sgRNA-2A-mCherry | Knuckles et al (2017) | |
pRRE200 homologous recombination reporter | Flemr & Buhler (2015) | |
pRRP200 homologous recombination reporter | Flemr & Buhler (2015) | |
pBluescript II KS(–) | Stratagene | |
psiCHECK-2 | Promega | C8021 |
Oligonucleotides are listed in Table S5 | IDT |