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Synthesis of Cy5/Cy3-labeled cDNA from 5 µg DNA-free total RNA per labeled cDNA was performed using CyScribe First-Strand cDNA Synthesis Kit (GE Healthcare, Freiburg, Germany) with enclosed random nonamer primers and Cy3- and Cy5-dUTP (GE Healthcare, Freiburg, Germany) following the manufacturers instructions.
High power microscopy (×400) was used to count migrant cells in 9 randomly chosen fields per labeled filter.
Note, these means correspond to the number of fluorophores per labeled particle, not the average number of fluorophores per particle over the entire sample.
However, if successive rounds of stem cell divisions take place after the initial labeling, then a dilution of labeling occurs each time, yielding new generations of neurons with less labeling, that is, fewer exposed silver grains per labeled neuron [ 12, 48].
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Customers are charged either a per label fee or a flat fee for a given number of shipments.
An amount of 20 μg of 3 was used per labelling.
Three treatments per labeling experiment were established, with each treatment prepared as described for our inoculation experiments.
The resulting index represents the number of cervical collateral per millimetre per labelled smHL CST fibres.
Data are normalised using 1/median ratio value for each identified protein group per labelled sample.
A limiting factor of cDNA microarray technology is the need for a substantial amount of RNA per labeling reaction.
In addition to reporting a per-worm accuracy, a per-label accuracy is provided.
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