Specific primers were designed for genes of interest retrieved from the transcriptome resources of M. fuliginosus, and fragments were amplified from either mixed stage or stage-specific cDNA. The PCR fragments were inserted into pGEM-T-easy vector (Promega) and cloned in Top10 E. coli cells (Invitrogen). DIG/FITC labeled RNA probes were generated by DIG RNA labeling mix (Roche) or using transcription reagents along with DIG-UTP/FITC-UTP (Roche) by SP6 or T7 polymerases (Roche).
Dig utp fitc utp
Manufactured by Roche
DIG-UTP/FITC-UTP are nucleotide analogs that can be used to label and detect nucleic acid sequences in various molecular biology applications. DIG-UTP is digoxigenin-labeled uridine triphosphate, and FITC-UTP is fluorescein isothiocyanate-labeled uridine triphosphate. These labeled nucleotides can be incorporated into nucleic acid probes or targets, enabling visualization and detection through immunodetection or fluorescence-based methods.
Automatically generated - may contain errors
2 protocols using dig utp fitc utp
1
Molecular Cloning and Probe Synthesis
2
Transcriptome Profiling via Cloning and In Situ Hybridization
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Gene transcripts were retrieved from the assembled transcriptome by bidirectional blast. Speci c primers were designed and fragments were ampli ed from either mixed stage or stage speci c cDNA. The PCR fragments were inserted into pGEM-T-easy vector (Promega) and cloned in Top10 E. coli cells (Invitrogen). DIG/FITC labeled RNA probes were generated by DIG RNA labeling mix (Roche) or using transcription reagents along with DIG-UTP/FITC-UTP (Roche) by SP6 or T7 polymerases (Roche). Gene orthology was inferred by reciprocal blast against GenBank.
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