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Real-Time Quantitative PCR:

Applications include gene expression analysis, allelic discrimination (e.g. SNP detection), pathogen detection and quantitation, mutation screening, and High Resolution Melting genotyping (Roche). For high-thoughput qPCR and digital PCR we have a Fluidigm BioMark Genetic Analysis System.

Instruments:

Applied Biosystems PRISM 7900HT Sequence Detection System (TaqMan)
Roche LightCycler 480 Real-Time PCR System

Both instruments have 96 and 384 well optical-plate capability. The optical-plates are instrument specific. Please check that you are using the correct plate for the instrument you choose. Only the Roche brand of 96-well plates may be used in the LightCycler. The ABI PRISM 7900HT will accept microfluidic card assays. Please note that we do not have the "FAST" capabilities for the ABI PRISM 7900HT.

Cost:

  • $40.00 per plate
  • Charges are incurred for missed appointments.

Using Our Services:

  • Reserve an appointment via our online WEBSEQ sign-up form.
  • We need a UCLA re-charge form to a create a WEBSEQ account for your lab if it does not already have one.
  • Before the appointment time, the customer brings the prepared sealed plate, along with the assay conditions for their run to the core. We need to know the dye-quencher combination for your probe, the PCR conditions you want, and the volume in the wells. We have forms you can use to supply this information. Please print and fill-out either the 96-well run information sheet or the 384-well run information sheet.
  • Core personnel set-up the run and post the results on WEBSEQ. We are located in CHS room 36-125. If you have any questions please contact us at webseq@genetics.ucla.edu.

Background Information:

Practical Information:

Below are potential resources for real-time qPCR assay design. These lists are by no means exhaustive.

Primer/Probe Design Sites


If you would like help in designing Taqman Fluorescent probes, send us the sequence of the region in the gene where the primers should be created. The region of interest should be between 150-250 base pairs in length. Send via email to

Uma Dandekar, GenoSeq Core Assistant Director
Email: uma@ucla.edu
Phone: 310-267-2461
Fax: 310-794-5446 OR webseq@genetics.ucla.edu as a Word document attachment.

Once we have emailed you the sequences for primers/probes sequences, order them from the vendor of your choice. A few such vendors are listed below.



If are using a SYBR Green mix and will be designing your own primers then please use a primer designer software program such as those listed below.


Please call us at 310-267-2461 if you have questions. Or contact:

Uma Dandekar, GenoSeq Core Assistant Director
Email: uma@ucla.edu
Phone: 310-267-2461
Fax: 310-794-5446
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