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In situ RT-PCR (Reverse Transcription) – zmodyfikowana wersja reakcji PCR, w której mRNA zostaje przepisane na cDNA za pomocą enzymu odwrotnej transkryptazy, następnie cDNA jest powielane przy użyciu starterów i z udziałem polimerazy.
In situ polymerase chain reaction (PCR) is a very powerful tool, which enhances our ability to detect minute quantities of a rare, single copy number, target nucleic acid sequences in freshly frozen or paraffin-embedded intact cells or tissue sections (1–10)....
1 lis 2007 · In this protocol we describe the in situ PCR method for the amplification of both DNA and mRNA targets [in situ reverse transcriptase-PCR (RT-PCR)], from frozen or paraffin-fixed tissue...
18 wrz 2014 · The protocol includes optimised methods for: (i) fixing, embedding, and sectioning of plant tissue; (ii) DNase treatment; (iii) in situ RT-PCR with the incorporation of DIG-labelled nucleotides; (iv) signal detection using colourimetric alkaline phosphatase substrates; and (v) mounting and microscopy.
This chapter describes the application of in situ PCR for neuropeptide mRNA localization. We provide here a detailed protocol for direct in situ reverse transcription (RT) PCR (RT-PCR) with nonradioactive probes after fixation and paraffin embedding or cryosectioning.
1 sty 2004 · The theory behind RT in situ PCR. It must be emphasized that one cannot perform target specific direct incorporation of the reporter molecule (e.g. digoxigenin dUTP) for DNA targets in paraffin embedded tissue sections which is referred to as in situ PCR.
In this protocol we describe the in situ PCR method for the amplification of both DNA and mRNA targets [in situ reverse transcriptase-PCR (RT-PCR)], from frozen or paraffin-fixed tissue sections, cell culture or other single-cell suspensions. Detection of amplicons can be achieved by the hybridization and detection of labeled probes.
