Total RNA was loaded onto agarose gels, separated by electrophoresis, and transferred to Gene Screen Plus membranes. Membranes were prehybridized in rapid hybridization buffer (Amersham, Arlington Heights, IL) and then incubated overnight at 68°C in hybridization buffer containing [32P]DNA probes (1 × 108 cpm) for Nrf2 or 18S mRNA [24 (link),25 (link),35 (link)]. Following hybridization and high stringency washing, membranes were exposed to X-ray film, and mRNA expression quantified by densitometry and normalized with respect to 18 S rRNA.
Rapid hybridization buffer
Manufactured by Cytiva
Rapid hybridization buffer is a buffer solution designed to facilitate efficient and rapid hybridization of nucleic acid probes during molecular biology techniques. It is optimized to provide the necessary ionic conditions and environment for effective hybridization, enabling faster and more reliable results.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using rapid hybridization buffer
1
Northern Blot Analysis of Nrf2 mRNA
2
Northern blot analysis of HO-1 expression
3
RNA Extraction and Northern Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rice leaves (1 g) were ground to a fine powder in liquid nitrogen and immersed in 5 mL extraction buffer (50 mM Tris-HCl pH 8, 300 mM NaCl, 5 mM EDTA (ethylenediaminetetraacetic acid), 2% (w/v) SDS (sodium dodecyl sulfate), 10 mM β-mercaptoethanol and 1 mM aurin tricarboxylic acid). Total RNA was precipitated by LiCl [51 (link)]. Afterward, total RNA (40 µg) samples were separated by 1% agarose/formaldehyde gel electrophoresis and then blotted onto Hybond-N+ nylon membranes (Amersham/GE Healthcare, Chicago, IL, USA). Northern hybridization was carried out in a rapid hybridization buffer (Amersham). Membranes were washed under high stringency washing conditions in 0.1× SSC (standard saline citrate) and 0.1% SDS at 65 °C. A 251 bp OsY37 fragment from the pGEM200 vector was radiolabeled with [α-32P]-dCTP (50 µCi) and used as a probe. Alternatively, for RT-PCR analysis, RNA was extracted from 100mg of green or senescing leaves using RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany), then quantified fluorometrically using Qubit RNA BR Assay Kit (ThermoFisher Scientific, Waltham, MA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!