Five micrograms of total RNAs were separated on 1% agarose gel containing 2% formaldehyde, and gels were treated with 0.05 N NaOH for 20 min to enhance transfer of high molecular weight RNAs, which step was essential to detect mNeat1_2/nNEAT1_2. RNAs were transferred on a positively charged Nylon membrane (Merck #11209299001) using standard protocol, and probe hybridization and detection were performed using DIG Easy Hyb (Merck #11603558001), anti-Dig AP (Merck #11093274910), and CSPD-Star (Merck #11685627001) following manufacturer's instructions. Chemiluminescence signals were detected with the ChemiDoc Touch Imaging System (Bio-Rad).
Dig easy hyb
Manufactured by Merck Group
DIG Easy Hyb is a laboratory equipment product from Merck Group. It is designed for hybridization of nucleic acid samples in a simple and efficient manner. The core function of DIG Easy Hyb is to facilitate the hybridization process, which is a crucial step in various molecular biology techniques.
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Lab products found in correlation
3 protocols using dig easy hyb
1
Northern Blot for lncRNA Detection
2
Sensitive Neat1_2 Transcript Detection
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Five micrograms of total RNA was separated on a denaturing gel (1% agarose gel, 1× MOPS, 2% formaldehyde), the gels were rinsed twice in distilled water for 15 min, and the RNA samples were hydrolyzed in 0.05 N NaOH for 20 min. This alkali treatment step is critical for detecting Neat1_2 transcripts. After the samples were soaked in water and subsequently incubated twice in 20× SSC for 45 min, the RNA samples were transferred to a positively charged nylon membrane (Merck, #11209299001) and were hybridized with probes at a concentration of 1 µg/mL in DIG Easy Hyb (Merck, #11603558001). The hybridized probes were detected with anti-Dig AP (Merck, #11093274910) and CDP-Star (Merck, #11685627001) following the manufacturer's instructions.
3
High-Molecular-Weight RNA Detection
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5 µg of total RNAs were separated on 1% agarose gel containing 2% formaldehyde, and gels were treated with 0.05N NaOH for 20 minutes to enhance transfer of high molecular weight RNAs, which step was essential to detect mNeat1_2/nNEAT1_2. RNAs were transferred on a positively charged Nylon membrane (Merck #11209299001) using standard protocol, and probe hybridization and detection were performed using DIG Easy hyb (Merck #11603558001),
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