By Hara N.
We supply a short new evidence of Wahl's vanishing theorem for rational floor singularities, utilizing major attribute process. Our procedure additionally provides a criterion for F-rationality in size 2
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Extra info for A characteristic p proof of Wahl's vanishing theorem for rational surface singularities
4. Inactivate the micrococcal nuclease activity by adding EGTA to a final concentration of 3 mM to the digest. 2. Replicase Assay 1. 0 µg viral mRNA; RNase-free distilled water, to bring the reaction mix to a total volume 20 µL. 2. Incubate the mixture at 32°C for 2–3 h. Virus Replication 51 Fig. 2. Replicase activity associated with untreated and micrococcal nuclease-treated open cores. The exogenous RNA added to some reaction mixtures (lanes 2 and 4) was produced in vitro by T7 transcription of a linearized plasmid containing a cDNA of the SA11 gene encoding NSP2 (gene 8).
3. Incubate the transcription mixture for 8–18 h at 37°C. 4. Treat with RNase-free DNase to remove the DNA template. 5. Deproteinize by extracting sequentially with phenol:chloroform and chloroform. 6. 5 vol of ethanol. Fig. 3. (continued) concentrations of NaCl (100–200 mM). (Third panel) Replicase assays contained 32P-UTP, open cores, no mRNA, or gene 8 mRNA and the indicated level of NaCl. The synthesis of 32P-labeled dsRNA in the reaction mixtures was quantitated by SDS-12% PAGE, followed by phosphorimaging.
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