Dot blot assays were conducted following standard protocols (Brown, 2001 ). Briefly, 3 μg of DNA from post‐autogamous cultures were blotted onto a nylon membrane (Hybond N+XL membrane, Amersham). Probes specific to Sardine and Thon transposons and actin (first 240 bp of the gene) labeled with α‐32P dATP (3,000 Ci/mmol) using RadPrime DNA Labeling System (Invitrogen) were used. The signal was quantified with ImageJ 1.48e.
Hybond n xl membrane
Manufactured by Cytiva
Hybond N+XL is a nylon membrane used for nucleic acid transfer and immobilization in Southern blotting and Northern blotting applications. It provides high binding capacity and excellent signal-to-noise ratio.
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Lab products found in correlation
2 protocols using hybond n xl membrane
1
Dot Blot Assay for Transposon Detection
2
RNA Expression Analysis via Northern Blotting
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Ten microgram of RNA were run in a 1.2% agarose denaturing gel and transferred to a nylon membrane (Hybond N+XL membrane, Amersham) by capillary blotting. After transfer, the membrane was crosslinked twice with UV (120,000 μJ/cm2). Specific probes labeled with α‐32P dATP (3,000 Ci/mmol) using RadPrime DNA Labeling System (Invitrogen) for ISWI1, ISWI5, and rRNA were used for hybridization. Membranes were screened using the Typhoon Imaging system (GE Healthcare).
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