... that gave a small amount of a product about 39 nt shorter than the template (Fig 4A) The relative quantity of this short product was variable in different experiments In all cases, slower migrating ... ATP and [32P]UTP The reaction mixture was further incubated at 25 °C for h [32P]RNA products were quantified after TCA precipitation of an aliquot of the reaction mixture as described in Experimental ... 10% (v ⁄ v) TCA The radioactivity incorporated into newly synthesized RNA was then determined To quantify and analyze the 32P-labeled RNA, the synthesis was stopped by adding 6.25 mm EDTA, 10 mm...
... animals (Fig 4) The CHH gene silencing was also corroborated by northern blot analysis and by semiquantitative RT-PCR The shrimps were killed 24 h after the injections, and the eyestalks were removed ... band in the all eyestalk total RNA sample tested (Fig 5B) Similar results were observed in the semiquantitative RT-PCR assays, which showed a defined DNA fragment of 216 bp, corresponding to CHH cDNA, ... 2006 FEBS J M Lugo et al A B Fig CHH gene silencing detection 24 h after the treatments by semiquantitative RT-PCR assays (A) PCR reaction with L schmitti CHH specific primer (B) PCR reaction...
... was used as template and newly synthesized RNA products were quantified by real-time quantitative RT–PCR (TaqMan assay), which allowed quantification of the starting copies of the reaction, rather ... Zhou, J., Pan, Z., Chen, L., Li, T & Guo, C (2003) Development of a fluorogenic quantitative PCR assay for rapid quantification of hog cholera lapinized virus Virologica Sin 18, 124–128 25 Yamashita, ... fluorogenic probe was designed with the following sequence: 5¢-TACAGGACAGTCGTCAGTAG TTCGACGTGA-3¢ RNA quantification was performed as follows: the precipitated RNA was reverse transcribed to cDNA using...
... accurate quantification of m R N A abundances problematic In contrast, the a R N A amplification technique amplifies m R N A in a linear fashion, using T7 R N A polymerase, and permits direct quantification ... enough cDNA to make a representative library The total double-strand cDNA quantity is determined simply by multiplying by the quantity of second-strand cDNA Keep a record of the counts per minute ... permits direct quantification of the relative abundances of individual m R N A species The ability to quantitate m R N A levels is critical, because expression analysis of multiple genes simultaneously...
... PAGE, subsequent Coomassie blue staining (to visualize the bacterially expressed proteins) and quantification of radioactive signals using BioImage analyzer BAS-1000 and the software TINA version ... separated on SDS-PAGE and analyzed using a BioImager Binding of NP to GST-NP was set to 100% (E) Quantification of separate experiments as shown under (D) (F) Intracellular distribution of NP and ... complexes were precipitated with glutathione-sepharose and the amounts of precipitated NP mutants were quantified by BioImaging (Figs 4C and 4D) NP mutants containing the N-terminal amino acids 1–330,...
... electrophoresis, and the products were visualized by autoradiography The reaction products were quantitated with a Phosphorimager (Molecular Dynamics Storm 800) by measuring the amount the amount...
... PAGE, subsequent Coomassie blue staining (to visualize the bacterially expressed proteins) and quantification of radioactive signals using BioImage analyzer BAS-1000 and the software TINA version ... separated on SDS-PAGE and analyzed using a BioImager Binding of NP to GST-NP was set to 100% (E) Quantification of separate experiments as shown under (D) (F) Intracellular distribution of NP and ... complexes were precipitated with glutathione-sepharose and the amounts of precipitated NP mutants were quantified by BioImaging (Figs 4C and 4D) NP mutants containing the N-terminal amino acids 1–330,...
... unsuitable for rapid clonal propagation The use of cuttings from young plants is limited because the quantity of cutting material which is produced by young ortet is low The rooting ability of older...