After treatment with EZ-Link HPDP-Biotin (Thermo Scientific), 500 ng of heat denatured (for 10 minutes at 65°C) total RNA were spotted on an Amersham Hybond-N+ positively charged nylon membrane (Cytiva). The RNA was ultraviolet-cross-linked to the membrane with Spectrolinker XL-1500 (Spectroline, ‘optimal crosslink’) and blocked for 10 minutes in blocking solution (PBS pH 7.5, 10% SDS, 1 mM EDTA). The membrane was then incubated in 1:1000 dilution of streptavidin-HRP (Pierce) in blocking buffer for 15 minutes. After washings with decreased concentration of SDS, the signal was recorded (Amersham ECL Western Blotting Detection Kit, Cytiva).
Amersham hybond n positively charged nylon membrane
Manufactured by Cytiva
Amersham Hybond-N+ is a positively charged nylon membrane used in nucleic acid transfer and immobilization techniques, such as northern, southern, and slot/dot blotting. It provides efficient and uniform binding of nucleic acids for downstream analysis and detection.
Automatically generated - may contain errors
2 protocols using amersham hybond n positively charged nylon membrane
1
RNA Labeling and Detection Assay
2
RNA Labeling and Detection Assay
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