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. 2021 Sep 27;2(4):100846. doi: 10.1016/j.xpro.2021.100846
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies

Biotin mouse lineage panel, diluted 1:40∗ BD Biosciences Cat# 559971
RRID: AB_10053179
PE streptavidin; diluted 1:100 BD Biosciences Cat# 554061
RRID: 554061
PE-Cy7 rat anti-mouse Ly-6A/E (Sca1) (clone D7); diluted 1:40∗ BD Biosciences Cat# 558162; RRID:AB_647253
APC anti-mouse CD117 (c-kit) (clone ACK2); diluted 1:40∗ BioLegend Cat# 135108; RRID:AB_2028407
Pacific Blue anti-mouse CD150 (clone TC15-12F12.2); diluted 1:40∗ BioLegend Cat# 115924; RRID:AB_2270307
APC-Cy7 anti-mouse CD48 (clone HM48-1); diluted 1:40∗ Thermo Fisher Scientific Cat# 47-0481-82; RRID:AB_2573962
Rabbit anti-human/mouse phospho-histone H2A.X (Ser139) (20E3); diluted 1:1000 Cell Signaling Technology Cat# 9718S; RRID:AB_2118009
Donkey anti-Rabbit IgG, Alexa Fluor 488; diluted 1:500 Thermo Fisher Scientific Cat# A21206;
RRID:AB_2535792

Biological samples

Human whole bone marrow and blood from healthy donors Lonza Cat# 1M-105

Chemicals, peptides, and recombinant proteins

FLT3-ligand, human PeproTech Cat# 300-19
IL-6, human PeproTech Cat# 200-06
SCF, human PeproTech Cat# 300-07
TPO, human PeproTech Cat# 300-18
SCF, mouse∗ PeproTech Cat# 250-03-10UG
TPO, mouse∗ PeproTech Cat# 315-14-10UG
Mitomycin C Sigma-Aldrich Cat# M0503
CAS:50-07-7
Triton X-100 Sigma-Aldrich Cat# T8787
CAS:9002-93-1
Sodium hydroxide (NaOH) pellets VWR Life Science Cat#0583-500G
CAS: 1310-73-2
Bovine serum albumin (BSA) Sigma-Aldrich Cat# A9647
CAS: 9048-46-8
Ammonium chloride solution STEMCELL Technologies Cat# 07800
CAS: 12125-02-9
EDTA (0.5 M) pH 8 Boston BioProducts Cat# BM150
CAS: 6381-92-6
Methanol Fisher Scientific Cat# A412-4
CAS: 67-56-1
70% Ethanol Fisher Scientific Cat# BP82011
CAS: 64-17-5
16% Paraformaldehyde (formaldehyde) aqueous solution Electron Microscopy Sciences Cat# 15710
CAS: 50-00-0

Experimental models: organisms/strains

Fancd2-/- and WT mice. C57BL/6 background, 2–6-month-old. Any gender. Parmar et al., 2009 N/A

Software and algorithms

FlowJo, version 10.5.3 N/A https://www.flowjo.com/
OpenComet plugin N/A http://www.cometbio.org/
ImageJ N/A http://fiji.sc

Other

EasySep Kit, human STEMCELL Technologies Cat# 19356
EasySep Buffer STEMCELL Technologies Cat# 20144
EasySep Magnet STEMCELL Technologies Cat# 18000
MidiMACS Separator Miltenyi Cat# 130-042-302
MACS MultiStand Miltenyi Cat# 130-042-303
CD34 MicroBead Kit, human Miltenyi Cat# 130-046-702
FcR Blocking Reagent, human Miltenyi Cat# 130-059-901
Lineage Cell Depletion Kit, mouse∗ Miltenyi Cat# 130-090-858
LS Column Miltenyi Miltenyi Cat# 130-042-401
MS Column Miltenyi Miltenyi Cat# 130-042-201
StemSpan SFEM II STEMCELL Technologies Cat# 09655
MethoCult H4434 Classic STEMCELL Technologies Cat# 04434
Methocult GF M3434 methylcellulose∗ STEMCELL Technologies Cat# 03444
PBS pH 7.4 Gibco Cat# 100-10-023
SYBR-Green dye Invitrogen Cat# S7567
Fetal bovine serum (FBS) Sigma-Aldrich Cat# F2442
Hanks balanced salt solution (HBSS) Lonza Cat# 10-547F
HEPES Buffer Fisher Scientific Cat# BP299-100
Penicillin-streptomycin (PenStrep) Gibco Cat# 15140-122
L-Glutamine Gibco Cat# 25030081
70 μm Cell strainer Fisher Scientific Cat# 087712
10 mL Eccentric tip syringe BD Cat# 305482
18G 1 Inch needle BD Cat# 305195
500 mL Bottle Top Vacuum Filter, 0.45 μm Corning Cat# 430514
CometAssay Single Cell Gel Electrophoresis Assay Kit Trevigen Cat# 4250-050-K
Normal goat serum (10%) Life Technologies Cat# 50062Z
SuperFrost Plus Slides Fisher Scientific Cat# 12-550-15
ProLong Gold Antifade Mountant with DAPI Life Technologies Cat# P36941
Double Cytofunnel. Disposable sample chambers with white filter cards Thermo Scientific Cat# 5991039
Round-Bottom Polystyrene Test Tubes with Cell Strainer Snap Cap, 5mL Fisher Scientific Cat# 08-771-23
SmartDish STEMCELL Technologies Cat# 27370
Non-tissue culture treated plate, 12 well flat bottom Falcon Cat# 351143
STEMvision STEMCELL Technologies Cat# 22000
Cytospin 3 Shandon Scientific Cat# 74000102

∗ Indicates reagents exclusive for mouse experiments