Total RNA was extracted from immature inflorescences (stages 1–12) as described in [50 (link)]. For small RNA blotting and detection, 10 to 12 μg were heated for 5 minutes at 95°C in 1,5 volume of standard formamide buffer and a constant volumes were loaded into a 15% Acrylamide (19:1 acrylamide:bis acrylamide), 8 M urea, 0,5X TBE gel and separated by electrophoresis. Samples were then electroblotted to Hybond-NX (GE Healthcare) and immobilized following a carbodiimide cross-linking procedure [87 (link)]. Hybridization was carried out in 15 ml of ULTRAhyb Buffer (Ambion) overnight at 50°C with a χ32P-ATP labelled probe (T4 polynucleotide kinase, Promega, 60 minutes at 37°C). Membranes were washed twice in 3X SSC, 5% SDS and once in 1X SSC, 1% SDS. Oligoprobe sequences are found in [50 (link)]. Acquisition and quantification of the signal was achieved with a PMI-FX (BioRAD) phosphoimager and the Quantity One software.
Pmi fx
Manufactured by Bio-Rad
The PMI-FX is a precision instrument designed for measuring the thermal conductivity properties of materials. It utilizes a steady-state heat flow technique to determine the thermal conductivity, thermal resistance, and thermal transmittance of a wide range of materials. The PMI-FX provides accurate and reliable measurements, enabling users to evaluate the thermal performance of their samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pmi fx
1
Small RNA Blotting and Detection
2
Quantifying Herbicide Uptake in Transgenic Plants
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V1 growth stage plants of event 125 of p35S::TDO and the control (six plants/entry) were sprayed with 1× nonradiolabeled mesotrione followed by applying 40 μg of 14C‐mesotrione droplets to the first trifoliate using a 0.5 μg μL−1 14C‐mesotrione solution in 1% of crop oil. Plants were pulled out of pots at 30 HAT and, after soil was washed from the roots, placed in cassettes, imaged and exposed to phosphoscreens. After 2–3 weeks of exposure in −80 °C, the phosphoscreens were scanned at 50 μm resolution using a Personal Molecular Imager (PMI)‐FX (Bio‐Rad).
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