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The fragments obtained corresponded to the IgRs to be analyzed and were then used as templates for a second PCR, which was carried out with the same pairs of primers as before, but in the presence of DIG DNA labeling mix (DNA labeling Mix, 10 × conc., Roche).
For triple labelings, NPY cRNA was labeled using Biotin-RNA labeling mix, 5-HT2C cRNA with Fluorescein-RNA labeling mix and 5-HT1A cRNA with DIG-RNA labeling mix.
For triple labelings, NPY cRNA was reacted using biotin-RNA labeling mix, 5-HT2C cRNA with fluorescein-RNA labeling mix and 5-HT1A cRNA with DIG-RNA labeling mix.
Briefly, sense and antisense DIG-labeled riboprobes were synthesized with a DIG RNA labeling mix according to the manufacturer's instructions (Roche, Basel, Switzerland).
The digoxigenin-labeled NtGSA sense and antisense riboprobes were synthesized using DIG RNA labeling Mix and T3, T7 RNA polymerase (Roche Diagnostics GmbH Mannheim, Germany), respectively.
Sections were hybridized with a DIG-labeled cRNA probe, which was synthesized using a DIG RNA labeling mix (Roche).
The DIG-labeled riboprobes were synthesized by T7 or SP6 polymerase with a DIG labeling mix (Roche) from a template containing the fragment from +4511 to +5159.
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Eight hundred and seventy-five pounds -- almost a half-ton -- of turkey filets were recalled from supermarkets in Arizona, California, Maryland, Ohio and Texas this week because of a big labeling mix-up.
The non-radioactive riboprobes were synthesized by in vitro transcription using T7 and SP6 polymerase (Promega), respectively, and were labeled with digoxigenin-labeling mix (Roche).
In situ hybridization was carried out as previously described [ 31] using the zfaanat2 probe [ 20] that was labeled with digoxigenin RNA-labeling mix (Roche).
Probes were transcribed using a digoxigenin-labeling mix (Roche).
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