Detection of Small Noncoding RNAs by In Situ Hybridization Using Probes of 2′-O-Methyl RNA + LNA
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Detection of Small Noncoding RNAs by In Situ Hybridization Using Probes of 2′-O-Methyl RNA + LNA. / Søe, Martin Jensen; Dufva, Martin; Holmstrøm, Kim.
In: Methods in Molecular Biology, Vol. 1173, 18.06.2014, p. 113-121.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Detection of Small Noncoding RNAs by In Situ Hybridization Using Probes of 2′-O-Methyl RNA + LNA
AU - Søe, Martin Jensen
AU - Dufva, Martin
AU - Holmstrøm, Kim
PY - 2014/6/18
Y1 - 2014/6/18
N2 - In situ hybridization is a powerful method to provide information about contextual distribution and cellular origin of nucleic acids, e.g., in formalin-fixed paraffin-embedded (FFPE) samples of tissue. Particularly the recently discovered classes of noncoding RNA (ncRNA) including endo-siRNAs and microRNAs require such a technique to enable their study and visualization in natural contexts, and in the last decade, many advances have been made, increasing our ability to specifically detect small ncRNAs. One of the key developments has been the demonstration of the superiority of using locked nucleic acid (LNA)-modified DNA probes for the detection of ncRNA in tissue. Here, we describe an alternative in situ hybridization protocol employing oligonucleotide probes consisting of combinations of LNA and 2´-O-methyl RNAs that under optimized hybridization buffer conditions can provide a highly sensitive assay performance with only 1 h hybridization time.
AB - In situ hybridization is a powerful method to provide information about contextual distribution and cellular origin of nucleic acids, e.g., in formalin-fixed paraffin-embedded (FFPE) samples of tissue. Particularly the recently discovered classes of noncoding RNA (ncRNA) including endo-siRNAs and microRNAs require such a technique to enable their study and visualization in natural contexts, and in the last decade, many advances have been made, increasing our ability to specifically detect small ncRNAs. One of the key developments has been the demonstration of the superiority of using locked nucleic acid (LNA)-modified DNA probes for the detection of ncRNA in tissue. Here, we describe an alternative in situ hybridization protocol employing oligonucleotide probes consisting of combinations of LNA and 2´-O-methyl RNAs that under optimized hybridization buffer conditions can provide a highly sensitive assay performance with only 1 h hybridization time.
KW - Faculty of Science
KW - Noncoding RNA
KW - MicroRNA
KW - LNA
KW - In Situ Hybridization
KW - 2'-O-methyl RNA
U2 - 10.1007/978-1-4939-0931-5_10
DO - 10.1007/978-1-4939-0931-5_10
M3 - Journal article
C2 - 24920364
VL - 1173
SP - 113
EP - 121
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -
ID: 117196737