Total RNA was purified from Xoc strain liquid cultures (OD600 = 1.0) using the EasyPure RNA Kit (Transgen Biotech). Total RNA was extracted from plant leaves using the MiniBEST Plant RNA Extraction Kit (TaKaRa). RNA (10-20 μg) was separated using 1% agarose gels containing 25 mM guanidinium thiocyanate, transferred to Hybond N+ nitrocellulose membranes (Merck Millipore), and cross-linked to membranes by UV radiation. Probes were 5′ labeled with digoxygenin. Membranes were prehybridized for 10 min at 42°C, and then incubated with labeled probes overnight. Membranes were then rinsed, dried, and visualized by phosphorimaging on the ChemiScope 3000 mini instrument as previously described (Wu et al., 2021 ).
Hybond n nitrocellulose membranes
Manufactured by Merck Group
Sourced in United States
Hybond N+ nitrocellulose membranes are highly absorbent sheets used for the transfer and immobilization of nucleic acids, proteins, and other biomolecules in various laboratory techniques. They provide a stable surface for the attachment of these biological samples, facilitating their detection and analysis.
Automatically generated - may contain errors
2 protocols using hybond n nitrocellulose membranes
1
RNA Extraction and Northern Blot Analysis
2
RNA Extraction and Northern Blot Analysis
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Total RNA was purified from Xoc BLS256 liquid cultures (OD600 = 1.0) using the EasyPure RNA Kit (Transgen Biotech, Beijing, China). RNA (10–20 μg) was separated in 1% agarose gels containing 25 mM guanidium thiocyanate, transferred to Hybond N+ nitrocellulose membranes (Merck Millipore, USA), and cross-linked to membranes by UV radiation. Probes were 5’-labeled with digoxygenin (DIG). Membranes were prehybridized for 10 min at 42°C, and then incubated with labeled probes overnight. Membranes were then rinsed, dried and visualized by phosphorimaging on a ChemiScope 3000 mini (CLiNX, Shanghai, China) as described previously [13 (link)].
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