By 姜 徳相
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Transfer the replicator to 95% ethanol. 6. Let excess ethanol drip off the pins, then flame. 7. Allow replicator to cool. 2. Robotic Pin Tools (BioMatrix Colony Arrayer System) Use the following procedure to clean and sterilize the replicator pins prior to use of the robot. 1. Fill the sonicator bath with 390 mL sterile distilled water. 2. Clean the replicator pins in the sonicator bath for 5 min. 3. Remove the water and fill the sonicator bath with 390 mL 70% ethanol. 4. Sterilize the replicator in the sonicator bath for 20 s per cycle, and repeat the cycle twice.
Control sample–Cy5). As a result, each mRNA sample will be reverse-transcribed twice. Aliquot equal amounts (1–2 mg) of control and experimental mRNA samples into microcentrifuge tubes. Dry down samples in a Speed-Vac (medium-heat setting to prevent RNA hydrolysis). 2. 5 ml DEPC water and 1 ml T18VN primer (1 mg/ml). 3. Denature the samples at 65°C for 5 min. 4. Incubate at 42°C for 5 min to anneal the T18VN primer. 5. 5 ml superscript II enzyme per sample. 5 ml of reaction mixture to each sample.
Our analyses show that in many cases, the differentially regulated genes correspond to physiological functions and known targets of well-characterized TFs. The expected binding sites of several TFs were also identified in the promoters of these genes. Moreover, novel DNA-binding sequences and putative targets were identified for less-characterized TFs. These results demonstrate that phenotypic activation is an effective approach to rapidly characterize TFs on a large scale, which should also be feasible in other organisms.