DBU means 1,8-diazabicyclo[5.4.0]undec-7-ene. It's a bulky base that permits elimination.
The first key step of the synthesis is the cycloaddition [4+2] of the singlet oxygen.
The data demonstrate greatly improved transcript integrity after extended duration RNA synthesis reactions using the HiScribe T7 High Yield RNA Synthesis Kit as compared to the competitor’s kit.
HiScribe™ T7 Quick High Yield RNA Synthesis Kit | NEB
We recommend evaluating the plasmid DNA template with the RNase Contamination Assay Kit (NEB #E3320) before plasmid DNA template is used in the HiScribe T7 High Yield RNA Synthesis Kit.
Speech Synthesis Markup Language (SSML) Version 1.1
RNA synthesized with the kit can be used for RNA structureand function studies, ribozyme biochemistry, as probes for RNase protectionassays and hybridization based blots, anti-sense RNA and RNAi experiments,microarray analysis and microinjection, as well as in vitro translation and RNAvaccines.
To synthesize high specific activity radioactive RNA probes or RNA with 100%substitution of one or more modified nucleotides we recommend using theT7 High Yield RNA synthesis kit (), in which the four nucleotides aresupplied separately.
DNA Template Preparation:
Linearized plasmid DNA, PCR products or synthetic DNA oligonucleotides canbe used as templates for in vitro transcription with the HiScribe T7 Quick High YieldRNA Synthesis Kit, provided that they contain a double-stranded T7 promoterregion upstream of the sequence to be transcribed.
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We recommend evaluating the plasmid DNA template with the RNase Contamination Assay Kit (NEB #E3320) before plasmid DNA template is used in the HiScribe T7 Quick High Yield RNA Synthesis Kit.
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For GC rich templates, or templates with secondary structures, incubation at 42°C may improve yield of full-length transcript.
If premature termination of transcription is found in high specific activity radiolabeled RNA probe synthesis, increase the concentration of “limiting NTP”.