PAN_qPCR

PAN: Protein and Nucleic Acid Facility

» Real-Time PCR

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• Sample Submission
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Genomic Services

• Next Gen Sequencing

• DNA Sequencing
& Fragment Analysis

• Single Cell Genomics

Protein Services

Located in rooms
B065 and B017

• Sample Submission Requirements

** Please check for the appropriate sample submission requirements for each service type before placing your orders

Full Service
Self Service

Full Service: (back to top)

• with Reverse Transcription: (Bioanalyzer QC included)

• Submission requirements:

• Total RNA

* Volume: at least 10ul per sample

• without Reverse Transcription:

• Submission requirements:

• cDNA

* Volume: ~18ul per triplicate

• Assays (TaqMan or SybrGreen)

* Volume: ~6ul per triplicate

• Recommended concentration ranges and QC values

Template	Concentration (F.C.)	260/280	260/230
RNA*	10ng - 2ug	2.0	2.0-2.2
cDNA**	1ng/ul - 10ng/ul	N/A	N/A
DNA*	1ng/ul - 10ng/ul	1.8	2.0-2.2
Primer/Assay	300nM - 900nM

*RNA/DNA must be measured on a Nanodrop and/or BioAnalyzer.
**Assume 100% conversion of RNA to cDNA and dilute RT accordingly.

Self Service: (back to top)

Currently the instrument is available for Self Service with the researcher providing all StepOnePlus compatable consumables and reagents. Please follow the instructions below to for reservation and usage information:

• Place an online order for "Run Only" under qPCR section at pan.stanford.edu.

• Book machine's time from our online scheduler. Please note that you may be required to create a log in for the scheduler, separate from the log in for the online ordering, since the two systems are currently not linked.

• Important Notes:

** First-time users must contact us for a free and quick tutorial on instrument and software before scheduling a run.

** Only use StepOnePlus compatible consumables and reagents.

§ ABI MicroAmp Fast Optical Plate (0.1ml) - Part #4346907.

§ ABI MicroAmp Optical Adhesive Film - Part #4360954

Free copy of the StepOnePlus software (PC only) is available through ABI/Life Technologies or through PAN.
Bring plate wrapped in foil. Do not place plate on ice, as ice may generate false flurescent signal. Plate is okay at RT for a short time. If plate needs to be stored for a few hours or a day you may store it in our fridge.
Bring plate layout and thermal cycling conditions for your SybrGreen or Taqman reagent with you.
A centrifuge is available in the lab for customer use.
After-hours access is available, please contact panpyro-qpcr@stanford.edu or Mike Eckart for access.

**Please note that the charge is per plate, not per hour, refer to the price sheet for detailed information.