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The cDNA was then used as a template for in vitro transcription reaction (BioArray High Yield RNA transcript labelling kit ENZO) using biotinylated CTP and UTP to produce labelled cRNA.
After second strand synthesis, the double-stranded cDNA template was used for in vitro transcription, in the presence of biotinylated nucleotides to produce labelled cRNA.
Radiolabelled toxin: Some researchers may grow toxigenic cyanobacteria in media with an added C source in order to produce labelled cyanotoxin for toxicokinetics studies or other research endeavours.
To produce labelled template for hybridisations aRNA was generated using a MessageAMP II aRNA Amplification Kit (Ambion) as per the manufacturer's instructions.
LMW RNA was isolated from these cells and used to produce labelled cDNA; these samples were then hybridized to the miRNA arrays.
Briefly, 25 μg of RNA was used to produce labelled cDNA by anchored oligo dT -primed reverse transcriptioligo dT -primedipt II reverse transcription (Life Technologies), in the presence of Cy3- or Cy5-dUTP (Amersham, Chalfont, St Giles, UK).
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"There's something very fake about this place, like it's trying so hard to be earthy and 'good' but it's really just some giant evil supermarket," said a caterer, who particularly dislikes Whole Foods' "conventional" versus "organic" produce labelling.
Biotin-11-UTP (Invitrogen™, AM8450) or [α-32P]-UTP (Perkin Elmer, NEG507T250UC) was added to the reaction as required to produce labeled RNA.
This effectively allows large datasets to be generated and used for classification, reducing the demand to manually produce labeled training instances.
Interestingly, human and rat brain extracts incubated with radiolabeled morphine produce labeled M3G and M6G [22], [22].
As remarks above, in contrast, standard tracers produce labeled neurons in the upper layers as well.
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