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 |