Quantitative Northern Blot Analysis

Northern blot analysis was performed as described previously (44 (link)). Briefly, total RNA (500 ng) was resolved by 12% PAGE containing 7 M urea, transferred to Hybond N⁺ membranes (GE Healthcare) and hybridized to 5΄-end labeled antisense probes (cyto tRNA^LysCUU: 5΄-GTCTCATGCTCTACCGACT-3΄; cyto tRNA^AlaAGC: 5΄-GCGCTCTACCACTGAGCTA-3΄; and mt tRNA^ValUAC: 5΄-GTGTTAAGCTACACTCTG-3΄). Typhoon 9400 and ImageQuant ver. 5.2 (GE Healthcare) were used for storage phosphor autoradiography visualization and quantification.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Shigematsu M., Honda S., Loher P., Telonis A.G., Rigoutsos I, & Kirino Y. (2017). YAMAT-seq: an efficient method for high-throughput sequencing of mature transfer RNAs. Nucleic Acids Research, 45(9), e70.

Publication 2017

Hybond N+ membranes Typhoon 9400 ImageQuant ver. 5.2

Autoradiography Membranes Northern blot analysis Phosphor Typhoon Urea

Corresponding Organization : Thomas Jefferson University

Top 5 similar protocols

Variable analysis

independent variables

Probe type (cyto tRNA^LysCUU, cyto tRNA^AlaAGC, mt tRNA^ValUAC)

dependent variables

Signal intensity (quantification) of the tRNA probes

control variables

Amount of total RNA (500 ng)
PAGE gel (12% containing 7 M urea)
Transfer to Hybond N+ membranes
Typhoon 9400 and ImageQuant ver. 5.2 for visualization and quantification

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!