Further Development 3.21: In Situ Hybridization

In situ hybridization. (A) Whole mount in situ hybridization for odd-skipped mRNA (blue) in a stage-9 Drosophila embryo. (B) Antisense RNA probe with uridine triphosphate conjugated to digoxigenin (DIG). (C) Illustration of two cells at the border of the odd-skipped expression pattern seen in the box in (A). The cell on the left is not expressing odd-skipped, whereas the cell on the right is. The antisense DIG-labeled RNA probe with complementarity to the odd-skipped gene becomes hybridized to any cell expressing odd-skipped transcripts. Following probe hybridization, samples are treated with anti-DIG antibodies conjugated to the enzyme alkaline phosphatase. When nitroblue tetrazolium chloride (NBT) and 5-Bromo-4-chloro-3-indolyl-phosphate (BCIP) are then added to the sample, alkaline phosphatase converts them to a blue precipitate. Only those cells expressing odd-skipped turn blue.

In situ hybridization. (A) Whole mount in situ hybridization for odd-skipped mRNA (blue) in a stage-9 Drosophila embryo. (B) Antisense RNA probe with uridine triphosphate conjugated to digoxigenin (DIG). (C) Illustration of two cells at the border of the odd-skipped expression pattern seen in the box in (A). The cell on the left is not expressing odd-skipped, whereas the cell on the right is. The antisense DIG-labeled RNA probe with complementarity to the odd-skipped gene becomes hybridized to any cell expressing odd-skipped transcripts. Following probe hybridization, samples are treated with anti-DIG antibodies conjugated to the enzyme alkaline phosphatase. When nitroblue tetrazolium chloride (NBT) and 5-Bromo-4-chloro-3-indolyl-phosphate (BCIP) are then added to the sample, alkaline phosphatase converts them to a blue precipitate. Only those cells expressing odd-skipped turn blue.

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