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Transcription of cards during in vitro growth of M. pneumoniae in SP-4 broth. A. Expression of cards along with other genes was analysed by DNA slot blot. M. pneumoniae <t>gene-specific</t> <t>PCR</t> products were blotted onto <t>Zeta</t> probe membranes. M. pneumoniae S1 cells were grown and harvested at early-log (24 h), mid-log (48 h), late-log (72 h) and stationary phases (120 h). 32 P-labelled cDNAs generated by reverse transcription of isolated total RNA were used as hybridization probes. Experiments were repeated three times. B. Expression of cards by relative real-time quantitative RT-PCR (qRT-PCR). M. pneumoniae S1 cells were grown and RNA isolated at specific growth phases. Real-time qRT-PCR was performed using SYBR green chemistry as detailed in Experimental procedures . Experiments were repeated two times. The average fold differences in expression levels of cards mRNA at various time points (when compared with the stationary phase) and standard deviations (SD) are presented.
Zeta Probe Membranes, supplied by BioDot Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Transcription of cards during in vitro growth of M. pneumoniae in SP-4 broth. A. Expression of cards along with other genes was analysed by DNA slot blot. M. pneumoniae gene-specific PCR products were blotted onto Zeta probe membranes. M. pneumoniae S1 cells were grown and harvested at early-log (24 h), mid-log (48 h), late-log (72 h) and stationary phases (120 h). 32 P-labelled cDNAs generated by reverse transcription of isolated total RNA were used as hybridization probes. Experiments were repeated three times. B. Expression of cards by relative real-time quantitative RT-PCR (qRT-PCR). M. pneumoniae S1 cells were grown and RNA isolated at specific growth phases. Real-time qRT-PCR was performed using SYBR green chemistry as detailed in Experimental procedures . Experiments were repeated two times. The average fold differences in expression levels of cards mRNA at various time points (when compared with the stationary phase) and standard deviations (SD) are presented.

Journal: Molecular Microbiology

Article Title: Mycoplasma pneumoniae Community Acquired Respiratory Distress Syndrome toxin expression reveals growth phase and infection-dependent regulation

doi: 10.1111/j.1365-2958.2010.07092.x

Figure Lengend Snippet: Transcription of cards during in vitro growth of M. pneumoniae in SP-4 broth. A. Expression of cards along with other genes was analysed by DNA slot blot. M. pneumoniae gene-specific PCR products were blotted onto Zeta probe membranes. M. pneumoniae S1 cells were grown and harvested at early-log (24 h), mid-log (48 h), late-log (72 h) and stationary phases (120 h). 32 P-labelled cDNAs generated by reverse transcription of isolated total RNA were used as hybridization probes. Experiments were repeated three times. B. Expression of cards by relative real-time quantitative RT-PCR (qRT-PCR). M. pneumoniae S1 cells were grown and RNA isolated at specific growth phases. Real-time qRT-PCR was performed using SYBR green chemistry as detailed in Experimental procedures . Experiments were repeated two times. The average fold differences in expression levels of cards mRNA at various time points (when compared with the stationary phase) and standard deviations (SD) are presented.

Article Snippet: All generated amplicons were purified by gel extraction, and 200 ng of each specific PCR product was blotted in triplicate on Zeta probe membranes by using Bio-Dot SF microfiltration apparatus as suggested by the manufacturer (Bio-Rad Laboratories, Hercules, CA).

Techniques: In Vitro, Expressing, Dot Blot, Generated, Reverse Transcription, Isolation, Hybridization, Quantitative RT-PCR, SYBR Green Assay