synthetic construct
Protocols: Synthetic DNA was generated by primer extension using template oligos (see methods section for sequences) Detailed protocol is available in methods section. Briefly, purified DNA was sonicated using a Covaris M220 system, to an average length of 450 bp. Sonicated DNA was treated with HIPS probe 1 (10 mM, 2 hr, RT) and biotinylated using the CuAAC reaction and biotin-PEG3-azide (2 hr, 37 °C). A custom P7 adapter was introduced onto DNA by ligation using a NEBNext Ultra II kit. Labelled DNA was isolated using magnetic streptavidin beads and unbound DNA was removed by washing. The enriched DNA was denatured on the beads using NaOH solution (RT) and then eluted using NaOH solution (70 °C). Eluted DNA was purified once again on magnetic streptavidin beads to remove any non-specific elution. A single primer extension was performed on eluted ssDNA, then a custom P5 adapter was introduced by ligation. Libraries were amplified by PCR.
ENA-FIRST-PUBLIC
2019-06-12T17:06:27Z
ENA-LAST-UPDATE
2018-09-04T14:35:12Z
External Id
SAMEA4883140
INSDC center name
Department of Chemistry University of Cambridge
INSDC first public
2019-06-12T17:06:27Z
INSDC last update
2018-09-04T14:35:12Z
INSDC status
public
Submitter Id
E-MTAB-7152:Synthetic_snAP
broker name
ArrayExpress
sample name
E-MTAB-7152:Synthetic_snAP
scientific_name
synthetic construct