TS No. 59: Cellular Localization of Transcripts via In situ RNA Hybridization

In situ hybridization is a widely used method that permits the localization of target mRNAs in a preserved tissue section. Most in situ hybridization methods use either non-isotopic or isotopically labelled riboprobes. The use of digoxigenin (DIG) compound is popular in the former type of labeling. For this, hybridized DIG-labelled ripoprobes are normally detected with high affinity anti-digoxigenin (anti-DIG) antibodies that are conjugated to alkaline phosphate that consequently allows for colorimetic visualization of the anti-DIG antibody conjugate using NBT (Nitro blue tetrazolium chloride)-BCIP (5-Bromo-4-chloro-3-indolyl phosphate, toluidine salt) substrate solution. There is also the alternative method of using fluorescence-labelled probes. In addition, various other in situ hybridization protocols have been developed and reported over the years.

Main Research: Dr Ooi Siew Eng