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. Author manuscript; available in PMC: 2021 Sep 7.
Published in final edited form as: Curr Biol. 2020 Jul 30;30(17):3484–3490.e4. doi: 10.1016/j.cub.2020.06.099

KEY RESOURCES TABLE

REAGENT or RESOURCE SOURCE IDENTIFIER
Antibodies
Anti-Digoxigenin-POD Fab fragment Sigma-Aldrich 11207733910
Bacterial and Virus Strains
N/A
Biological Samples
N/A
Chemicals, Peptides, and Recombinant Proteins
Agarose, Type II-A, Medium EEO Sigma-Aldrich A9918
Penicillin-Streptomycin 100x Gibco 15-140-122
TDO Inhibitor 680C91 Sigma-Aldrich SML0287
Dimethysulfoxide (DMSO) Sigma-Aldrich D8418
Diethyl pyrocarbonate Sigma-Aldrich D5758
Paraformaldehyde Electron microscopy sciences 15714
2X NLS Cas9 protein Synthego
Formamide Sigma 47670–2.5L-F
DIG wash and block set (Roche blocking buffer) Sigma 11585762001
RNA from yeast Sigma 10109223001
SP6 RNA polymerase Roche M0207L
DIG-labeling nucleotide mix Roche 11277073910
Fluoromount-G Southern biotech 0100–20
Ethanol Pharmco 111000200
Critical Commercial Assays
pGEM t Easy vector Promega A1360
Cy5 TSA plus kit Perkin Elmer NEL752001KT
Monarch DNA Kit NEB T3010L
Monarch Gel Extraction kit NEB T2010L
Deposited Data
TDO locus of Doryteuthis pealeii GenBank MT648678
Doryteuthis pealeii RNAseq data Bioproject PRJNA641326
Experimental Models: Cell Lines
N/A
Experimental Models: Organisms/Strains
Adult Doryteuthis pealeii Wild caught in the Vineyard Sound near Woods Hole
Oligonucleotides
In situ PCR primer 1 IDT GAGCAATCGCGTCAAGTACA
In situ PCR primer 2 IDT GGCGTTGTCTTCAGGGTAGA
sgRNA 1 PCR forward primer nested reaction 1 IDT GCCTCAAAACAACCCATATTATTGAGG
sgRNA 1 PCR reverse primer nested reaction 1 IDT GAGTTGTAGCGCATCTGAGCAC
sgRNA 1 PCR forward primer nested reaction 2 IDT TAAATACTTGTGTTCATAGGGTACAC
sgRNA 1 PCR reverse primer nested reaction 2 IDT GGTAAACCCGCTCTGAGTTATTTCC
sgRNA 2 PCR forward primer nested reaction 1 IDT GCGTGCTATTCTGCATTAGCAC
sgRNA 2 PCR reverse primer nested reaction 1 IDT CGTTAAACCAGTTCTGCCCTCAAG
sgRNA 2 PCR forward primer nested reaction 2 IDT CCCTAACCATAACCTTAACGTCTC
sgRNA 2 PCR reverse primer nested reaction 1 IDT GCATTCTGTACGATGACACTAAGC
CRISPR sgRNAs
sgRNA1 Synthego CAUCCAAUCAGUGCCGAAGC
sgRNA2 Synthego UGGCAGCUGAGGUUCGUGUU
Software and Algorithms
MUSCLE (39)
Fig tree Software http://tree.bio.ed.ac.uk/software/figtree
Fast tree 2 software (40) http://www.microbesonline.org/fasttree/
Star v. 2.7.0 (42)
Bowtie2 (44)
SAMtools Genome Research Limited http://www.htslib.org/
Indel.py This study https://github.com/pipedq/CRISPR_G0_Genotyping
Cumulative.py This study https://github.com/pipedq/CRISPR_G0_Genotyping
Event_frequency.py This study https://github.com/pipedq/CRISPR_G0_Genotyping
Other
Quartz glass capillaries for making injection needles Sutter Instruments QF100-70-10