Real Time and Digital PCR

Quantitative polymerase chain reaction (PCR) is a method by which the amount of the PCR product can be determined, in real-time, and is very useful for investigating gene expression. 

7900HT Fast Real-Time PCR System »

The 7900HT System by Applied Biosciences is a high-throughput real-time PCR system that detects and quantitates nucleic acid sequences. 

Fluidigm Biomark Genetic Analysis System »

The Biomark is an exciting system that melds Microfluidics and Real-time PCR.  The system enables 9216 reactions in 7nL volume allowing real-time profiling of 96 target genes against 96 samples in a single run.

Please use dnaLIMS to reserve time on the equipment. Contact BRF if you require training or any other assistance. Users must supply their own consumables, a range of which can be purchased using dnaLIMS and collected when you visit the lab.

General

1. The BRF operates on a cost recovery basis. The percentage of subsidization is determined by The John Curtin School of Medical Research.

2. No samples or orders will be processed without an authorised* sample submission/order form and valid charge code. (*Authorised by the signature of the PI/Lab Head, or by granting of electronic access to BRF ordering systems with the authority of the PI/Lab Head).

3. It is a condition of the contract between the ACRF and JCSMR/ANU that "the Foundation (ACRF) is to be acknowledged in all scientific publications which utilise the Facility (BRF) at the Institute (JCSMR)". All work performed in the BRF, whether full service or not, and the use of any BRF resources should be acknowledged in all publications arising from that work. This should be in the “Material and Methods” section of the paper (see examples below). Any further individual acknowledgements are solely at your discretion.

4. A reference to the publication should be sent to the Manager of the BRF once the paper is published (includes theses).

5. BRF collaborations and co-authorship: although the BRF is primarily a service unit, there are instances where BRF staff make significant contributions to either the technical or intellectual input of the project. This should be discussed with the staff member concerned prior to the commencement of the collaboration.

Consumables and data

  1. The customer agrees to cover the cost of any consumables ordered on their behalf if the customer fails to supply the correct quantity and quality of the starting material required for processing. Details of the quality and quantity required are on the BRF website and sample submission form.
  2. Customers are responsible for archiving data generated by the BRF. Data generated by the BRF are solely for the use of the customer and their collaborators. Data is not to be sold to a third party. The BRF shall not be responsible for data output generated from samples that deviate from recommended protocols as requested by the customer.


  3. Upon receipt of consumables and/or data, the customer accepts responsibility for the correct handling, use, storage and disposal of the consumables/data.
  4. The BRF extends no warranties of any kind in respect to the consumables.

  Any consumables sold may have hazardous properties.  The customer agrees to use appropriate caution and safeguards as not all properties are known.
  5. Consumables sold by the BRF shall not be transferred to another party without the written consent of the BRF.  The BRF is not responsible for any losses arising from the use of consumables. The BRF will not be liable to the customer for any loss, claim or demand made by the customer due to acceptance, handling, use, storage or disposal of consumables and/or data by the customer, except to the extent permitted by law when it is the result of willful misconduct on the part of the BRF or its employees.

Acknowledgement in Publications

Examples

Real-time PCR

“Amplifications were performed in 384-well optical reaction plates (Applied Biosystems) with a 7900HT Fast Real-Time PCR System at the Genome Discovery Unit - ACRF Biomolecular Resource Facility, The John Curtin School of Medical Research, Australian National University using SDS 2.4 software to analyse raw data.”

DNA Sanger Sequencing

Amplified PCR products were purified and sequenced on an AB 3730xl DNA Analyzer (at the Genome Discovery Unit - ACRF Biomolecular Resource Facility, The John Curtin School of Medical Research, Australian National University) following the manufacturer's protocol (Applied Biosystems 2002).”

Peptide synthesis

Peptides were synthesized chemically using the 9-fluorenylmethyloxycarbonyl (Fmoc) method on a CEM Microwave-assisted Peptide Synthesizer and purified by one round of C18 reversed-phase HPLC by the Genome Discovery Unit - ACRF Biomolecular Resource Facility at the John Curtin School of Medical Research, Australian National University. As required, the N- or C-terminus, or both, were protected by acetylation or amidation, respectively.”

Tetramer synthesis

Cells were surface stained with APC-labelled tetramers consisting of murine class I MHC molecule (H-2Db), b2-microglobulin and influenza nucleoprotein peptide NP366–374. Tetramers were synthesised at the Genome Discovery Unit - ACRF Biomolecular Resource Facility at The John Curtin School of Medical Research, Australian National University using BirA enzyme synthesized as described (O’Callaghan et al., 1999). [O’Callaghan, C.A., Byford, M.F., Wyer, J.R., Willcox, B.E., Jakobsen, B.K., McMichael, A.J. and Bell, J.I (1999). BirA Enzyme: Production and Application in the study of membrane receptor-ligand interactions by site-specific biotinylation.Anal. Biochem. 266, 9-15.]”