scholarly journals Demonstration of microRNA using in situ hybridisation on formalin fixed paraffin wax samples using conventional oligonucleotide probes: a comparison with the use of locked nucleic acid probes

2020 ◽  
Vol 77 (3) ◽  
pp. 135-141 ◽  
Author(s):  
A Warford ◽  
NS Rahman ◽  
DA Ribeiro ◽  
P Uysal Onganer
Keyword(s):  
Nucleic Acid ◽  
In Situ Hybridisation ◽  
Paraffin Wax ◽  
Locked Nucleic Acid ◽  
Oligonucleotide Probes ◽  
Nucleic Acid Probes ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed

Distinguishing L from M photopigment coding sequences by hybridization to novel locked nucleic acid (LNA) oligonucleotide probes

2008 ◽  
Vol 25 (3) ◽  
pp. 283-287
Author(s):  
CHRISTINA PETTAN-BREWER ◽  
LI FU ◽  
SAMIR S. DEEB
Keyword(s):  
Nucleic Acid ◽  
Locked Nucleic Acid ◽  
Macaca Nemestrina ◽  
Oligonucleotide Probes ◽  
Coding Sequences ◽  
Cone Mosaic ◽  
Target Nucleic Acid ◽  
High Degree

Many attempts have been made over the years to distinguish human and primate L (long-wavelength sensitive) from M (middle-wavelength sensitive) cone photoreceptors using either immunohistochemistry or in situ hybridization. These attempts have been unsuccessful due to the very high degree of identity between the sequences of the L and M proteins and encoding mRNAs. The recent development of chemically modified oligonucleotide probes, referred to as locked nucleic acid (LNA) probes, has shown that they hybridize with much greater affinity and specificity to the target nucleic acid. This has greatly increased the potential for differentiating L from M cones by in situ hybridization. We have designed LNA oligonucleotide probes that are complementary to either the L or M coding sequences located in exon 5 of the Macaca nemestrina L and M pigment genes. We have shown that the LNA-M and LNA-L probes hybridize specifically to their respective target nucleic acid sequences in vitro. This result strongly suggests that these probes would be instrumental in rapidly distinguishing L from M cone in the entire retina, and in defining the cone mosaic during development and in adults.

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