Quantitative Real-Time PCR
Plate Running Service
The Plate running service is intended only for user-prepared plates. To use the service, please provide your billing address, in order to create specific user name and password. The login details are required to access the results folder on our file server.
We currently operate QuantStudio™ 12K Flex Real-Time PCR System (Thermo Fisher Scientific).
If you wish to have a customized service or a full service (preparation of the samples included), please contact us at taqman(a)utu.fi.
If you bring the plate to us, below is a list of details you should consider.
- You have the right format
- We can run :
- 96-well plates (volume 100 µl, 4titude, 4ti-0912 or equivalent barcoded plate)
- 384-well plates (4titude,4ti-0384 or equivalent barcoded plate).
Both plate types and plate seals are available through our plastic ware storage. Please note that all plates coming to the plate running service need to be barcoded. For those users who do not have access to our common storage; you need to order by mail taqman (at) utu.fi what you want to purchase and we will then deliver your order to the parcel drop zone. 96 and 384 well plates are sold in packages of 10 and seals are sold in packages of 100.
- Prepare your plate, put it in a plastic (minigrip) bag, identify the bag with your username and bring it to the cold room “Kyösti” into a box “qPCR Plate Running Service” in Biocity, A-stairs, 5th floor (BTK).
- Make the run file (eds.file) for your plate with QuantStudio 12K analysis software (you can get the software from us free of charge).
- Thermo Fisher provides software options for data analysis:
- Thermo Fisher Cloud is the recommended way to view and analyze your run data. You can access the cloud here.
- QuantStudio 12 K Flex software (current version 1.2.2) is used with QuantStudio instrument for creating and analyzing plate run files. To get your copy of the QuantStudio software, please contact us at taqman (at) utu.fi.
- Send the EDS file as an e-mail attachment to taqman (at) utu.fi. All the 96 well plates which are submitted in the plate running service on weekdays before 1 pm will be placed on the running queue of the same day, and you will get the results during the next workday. The 384 plates are run from Mondays to Thursdays, during working hours, when there is suitable time to change the block.
You can have a look at the plate queue in here.
- If you would like to get the plate back, please mention it in the email.
Universal Probe Library
Choose your probes from Roche Universal Probe Library using their assay design center. The probes are available through us.
Roche Universal Probe Library contains short hydrolysis probes of only 8 or 9 bases. The high melting temperatures characteristic of longer probes are retained by using Locked Nucleic Acid (LNA) nucleotide chemistry in these shorter probes. Because the probes are only 8 or 9 bases long, each probe can hybridize to over 7,000 transcripts; thus, a set of only 165 probes can enable the quantification of virtually any transcript in a transcriptome. Assay specificity is attained by the combination of specific primers and the Universal Probe Library probe. More information are available here.
The Universal ProbeLibrary probes are compatible with the TaqMan technology. The probes are available in 125 µl original tubes and 10 µl aliquots. The concentration is 10 µM, which is diluted to a working concentration of 1 µM by adding 90 µl of nuclease-free water into a 10 µl aliquot. The final concentration in with standard PCR settings is 0,1 µM (i.e. 100 nM). Thus, one 10 µl aliquot is sufficient for 100 reactions, i.e. one 96-well plate.
You can also order housekeeping gene sets from us, either B-actin or GAPDH for human or for mouse / rat. One set includes 10 µl aliquots of primers (300 µM) and probe (200 µM, FAM as a reporter).
To order probes, please contact us at: taqman (at) utu.fi
When you order probes, please specify:
- the Universal Probe number(s)
- the amount of each you need
- your name and your research group
We will bring the probes to the “Kyllikki” freezer room in BTK (5th floor, room number 5074) where you can pick them up. The probes are delivered at the latest in three working days (usually sooner).