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. Author manuscript; available in PMC: 2018 Dec 21.
Published in final edited form as: Mol Cell. 2017 Dec 21;68(6):1095–1107.e5. doi: 10.1016/j.molcel.2017.11.032
REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Monoclonal ANTI-FLAG M2 antibody Sigma F9291-1MG
Anti-GAPDH antibody Abcam ab9485
Amersham ECL Mouse IgG, HRP-linked Ab (from sheep) GE Healthcare Life Sciences NXA931-1ml
Bacterial and Virus Strains
One Shot TOP10 chemically competent E. coli Life Technologies C404006
Biological Samples
DMEM VWR 45000-304
Opti-MEM Life Technologies 31985062
FBS Clontech 631367
Chemicals, Peptides, and Recombinant Proteins
[γ-32P]ATP PerkinElmer NEG035C001MC
[α-32P]UTP PerkinElmer NEG007H001MC
[α-32P]GTP PerkinElmer NEG006H500UC
Pronase EMD Millipore 537088-50KU
Notl-HF New England Biolabs R3189L
cOmplete, Mini, EDTA-free protease inhibitor cocktail tablets Sigma 11836170001
TRI Reagent solution Life Technologies AM9738
Phenol:chloroform:isoamyl alcohol 25:24:1 Sigma P2069-100ML
Chloroform J.T.Baker Analyical 9180-01
SUPERase-In Life Technologies AM2696
TURBO DNase Life Technologies AM2239
Yeast tRNA Life Technologies 15401011
Dynabeads MyOne Streptavidin C1 Life Technologies 65002
ANTI-FLAG M2 magnetic beads Sigma M8823
3X-FLAG peptide Sigma F4799-4MG
GE Healthcare Amersham Hybond-XL membranes Thermo Fisher Scientific 45001147
GE Healthcare Amersham Hybond-NX membrane VWR 95038-412
Whatman Protran nitrocellulose membrane Sigma Z670898
PVDF pre-cut blotting membrane, 0.2 μm pore size Life Technologies LC2002
GE Healthcare Amersham Hyperfilm ECL Thermo Fisher Scientific 45001504
GE Healthcare Amersham ECL prime western blotting detection reagent Thermo Fisher Scientific 45002401
Gel loading buffer II (denaturing PAGE) Life Technologies AM8547
Whatman Nytran SuPerCharge (SPC) TurboBlotter Sigma WHA10416302
Critical Commercial Assays
mMESSAGE mMACHINE SP6 Life Technologies AM1340
RNeasy Mini Kit QIAGEN 74104
QuikChange Lightning Multi Site-Directed Mutagenesis Agilent 210515
Phusion High-Fidelity DNA polymerase New England Biolabs M0530L
T4 PNK New England Biolabs 101228-172
MAXIscript T7 RNA polymerase Life Technologies AM1312
Vaccinia capping system New England Biolabs M2080S
Lipofectamine 2000 Life Technologies 11668019
NuPAGE Novex 4-12% Bis-Tris Gel 1.0 mm Life Technologies NP0321BOX
NuPAGE MOPS SDS running buffer Life Technologies NP0001
NuPAGE transfer buffer Life Technologies NP00061
Micro Bio-Spin P-30 gel columns Bio-Rad 7326250
NorthernMax-Gly kit Life Technologies AM1946
ULTRAhyb Ultrasensitive hybridization buffer Life Technologies AM8670
ULTRAhyb-Oligo hybridization buffer Life Technologies AM8663
Deposited Data
Raw data This paper, Mendeley Data https://data.mendeley.com/datasets/fjg3b788k5/draft?a=f466501d-4dac-4b86-8d57-4cecb1fc3376
Experimental Models: Cell Lines
Human: HEK293T ATCC CRL-3216
Experimental Models: Organisms/Strains
Zebrafish ZIRC AB
Oligonucleotides
See Table S1 This study Table S1
Recombinant DNA
pCS2+-eGFP This study N/A
pCS2+-drAgo2 This study N/A
pCS2+-FLAG3-drAgo2 This study N/A
pCS2+-FLAG3-drAgo2D–A This study N/A
pCS2+-FLAG3-drAgo2D–E This study N/A
pCS2+-FLAG3-drAgo2Y–F This study N/A
pCS2+-FLAG3-drAgo2DY–EF This study N/A
pCS2+-FLAG3-hsN-drAgo2 This study N/A
pCS2+-hsAgo2 This study N/A
pCS2+-FLAG3-hsAgo2 This study N/A
pCS2+-FLAG3-hsAgo2D–A This study N/A
pCS2+-FLAG3-hsAgo2E–D This study N/A
pCS2+-FLAG3-hsAgo2F–Y This study N/A
pCS2+-FLAG3-hsAgo2EF–DY This study N/A
pCS2+-FLAG3-drN-hsAgo2 This study N/A
pCS2+-Zeocin This study N/A
pCS2+-Zeocin-miR-430 This study N/A
pCS2+-Zeocin-miR-430-10–11 mm This study N/A
pCS2+-Zeocin-miR-430-G–G mm This study N/A
pMAX GFP Amaxa VDF-1012
pcDNA3-drAgo2 This study N/A
pcDNA3-FLAG3-drAgo2 This study N/A
pcDNA3-FLAG3-drAgo2D–E This study N/A
pcDNA3-FLAG3-drAgo2Y–F This study N/A
pcDNA3-FLAG3-drAgo2DY–EF This study N/A
pcDNA3-FLAG3-drAgo2D–A This study N/A
pcDNA3-hsAgo2 This study N/A
pcDNA3-FLAG3-hsAgo2 This study N/A
GFP-3xPTmiR-1 Giraldez lab N/A
Software and Algorithms
MATLAB Mathworks
Multi Gauge 2.2 Fujifilm
GraphPad Prism GraphPad Software
Other