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Notes: The authors of this study investigated the microbial mechanisms associated with the reduction of methane production and emission from rice fields observed with intermittent field drainage. They looked in particular at the abundance of mcrA gene copies and transcripts from rice paddy soil fauna. The mcrA gene encodes the alpha subunit of methyl coenzyme M reductase. 

Total nucleic acid was extracted from soil samples using a phenol-chloroform procedure. For RNA analyses, DNA was hydrolyzed using RQ1 RNase-free DNase in the presence of 0.2µl Recombinant RNasin® Ribonuclease Inhibitor and then further purified using a commercial kit. cDNA synthesis was carried out using the Improm-II™ Reverse Transcription System, again in the presence of 1.0µl Recombinant RNasin® Ribonuclease Inhibitor. A clone library of transcripts was generated using the pGEM®-T Easy Vector System. The transcript standard for quantitative mcrA analysis was prepared from the in vitro transcript of a mcrA clone using the Riboprobe® in vitro Transcription Systems. (4241)

Notes: The authors of this study investigated the function of the daf-15 gene in C. elegans. The daf-15 gene encodes the C. elegans ortholog of raptor, a protein involved in the TOR signaling pathway. In C. elegans, daf-15 transcription is regulated by DAF-16, a FOXO transcription factor which is itself regulated by the insulin/IGF-1 signaling pathway. This work links regulation of the TOR pathway, which controls cell growth, to the insulin/IGF-1 pathway, known to affect lifespan, development and metabolism. Three candidate daf-15 genes were amplified by PCR and cloned into the pGEM^®-T Vector. The Riboprobe^® SP6/T7 transcription system was used to transcribe RNA from the candidate clones. Semiquantitative RT-PCR was performed to compare daf-15 mRNA levels in daf-2 and daf-16; daf-2 mutant animals. The mRNA was purified from total worm RNA using the PolyATtract^® mRNA Isolation System. (3633)

Notes: Total RNA was isolated from Arabidopsis tissues using the RNAgents^® Total RNA Isolation System. The RNA was further processed into the poly(A) fraction using the PolyATtract^® mRNA Isolation System. The isolated RNA was used in Northern blot analysis. Blots were probed with a ³²P-labeled RNA probe generated from a cDNA cloned into the pGEM^®-T Easy Vector using the T7 Riboprobe^® in vitro Transcription System. Bioluminescence from transgenic plants containing a firefly luciferase reporter was visualized by spraying the plants with a solution of 5mM Beetle Luciferin, Potassium Salt in 0.1% Triton^® X-100. (2570)

Notes: The PolyATtract^® System 1000 was used to isolate poly(A)+ RNA directly from myxoma virus-infected RK13 rabbit kidney cells. To obtain late viral RNAs, the RNA was isolated from the cells 16 hours after infection. For the early viral RNAs, the cells were treated with cycloheximide for 10 hours after infection prior to RNA isolation. The isolated RNA was used for Northern blot analysis with RNA probes prepared using the Riboprobe^® System. (0966)

Notes: The Riboprobe^® in vitro Transcription System was used to produce RNA probes for Northern analysis. (0417)

Notes: Promega's Tth DNA Polymerase, pGEM^®-T Vector System and Riboprobe^® System were used in this study. (2002)