SAMN38227369
External Id
CIC bioGUNE
INSDC center name
2024-09-19T00:00:00Z
INSDC first public
2024-10-04T01:48:20.450Z
INSDC last update
SRS22815868
INSDC secondary accession
live
INSDC status
MIGS.ba
NCBI submission model
MIGS.ba.6.0
NCBI submission package
SRS22815868
SRA accession
not applicable
collection_date
Keywords: GSC:MIxS;MIGS:6.0
description
PAM determination assay was performed incubating of 170 nM of -synCas-crRNA (targeting 23 nt downstream the 7 random nucleotides) and 100 nM of PCR fragment from the DNA library in 1 x NEB 3.1 buffer (50 mM Tris-HCl pH 7.9, 100 mM NaCl, 10 mM MgCl2, 100 g ml1 BSA) for 1 hour at 37 C. Reaction was stopped added 6X loading dye (NEB) with EDTA and run 2% agarose gel. Gels were dyed with SYBR gold (ThermoFisher) and imaged with ChemiDoc XRS + System (Bio-Rad). The longer fragment, that contained the 7 random nucleotides, was purified from the agarose gel with GeneJet Gel Extraction kit (ThermoFisher). Sequencing libraries of the purified fragments were prepared following Microplex-library-prep-v3 User Guide (version 2 15_06_2021) with the corresponding kit Microplex-library-prep-v3 (Diagenode, Cat. # C05010001) and indexes Diagenode 24 Dual indexes for MicroPlex Kit v3 (Diagenode, Cat. # C05010003).
design_description
not applicable
env_broad_scale
not applicable
env_local_scale
not applicable
env_medium
S_02_1.fastq.gz
filename
S_02_2.fastq.gz
filename2
fastq
filetype
missing
geo_loc_name
Illumina NovaSeq 6001
instrument_model
not applicable
isol_growth_condt
not applicable
lat_lon
S_02
library_id
paired
library_layout
other
library_selection
SYNTHETIC
library_source
CLONE
library_strategy
not applicable
num_replicons
Escherichia coli
http://purl.obolibrary.org/obo/NCBITaxon_562
organism
ILLUMINA
platform
not applicable
ref_biomaterial
missing
strain
alpha-synCas
title
MIGS Cultured Bacterial/Archaeal sample from Escherichia coli
title