Real Time qPCR System


Applied Biosystems

7900HT Fast Real-Time System with Power SYBR Green chemistry
In molecular biology, real-time polymerase chain reaction, also called quantitative real time polymerase chain reaction (Q-PCR/qPCR), is a laboratory technique based on the polymerase chain reaction, which is used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of a specific sequence in a DNA sample. The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is quantified as it accumulates in the reaction in real time after each amplification cycle. SYBR Green Chemisty intercalates with double-stranded DNA. Frequently, real-time polymerase chain reaction is combined with reverse transcription to quantify messenger RNA (mRNA) in cells or tissues.

SBI System Biosciences

miRNome micro RNA Profiling
In genetics, microRNAs (miRNA) are single-stranded RNA molecules of 21-23 nucleotides in length, which regulate gene expression. miRNAs are encoded by genes from whose DNA they are transcribed but miRNAs are not translated into protein (i.e. they are non-coding RNAs); instead each primary transcript (a pri-miRNA) is processed into a short stem-loop structure called a pre-miRNA and finally into a functional miRNA. Mature miRNA molecules are partially complementary to one or more messenger RNA (mRNA) molecules, and their main function is to down-regulate gene expression. They were first described in 1993 by Lee and colleagues in the Victor Ambros lab, yet the term microRNA was only introduced in 2001 in a set of three articles in Science.

To Run qPCR Samples Through The Genomics Core:

  • Step 1: Obtain 96 well full-skirted PCR plate from the genomics core to prepare your samples in.

  • Step 2: Download qPCR Request Form excel file and follow the instructions on the form to prepare your plate.

  • Step 4: Drop off your PCR plate containing cDNA + Primers in a total volume of 17uL per well to Genomics Core.

  • Step 5: The Core will e-mail you with the results containing: dissociation curves, amplification curves, and results for all samples/primer groups.