Tuesday, September 8, 2020

QuantStudio™ 6 Pro and 7 Pro Real-Time PCR Systems

Today is my first day in the lab!

Here are some of the things I see in the lab:
Frog fish https://www.bing.com/videos/search?q=frog+fish+eats&docid=608039039962123360&mid=118E8987D1FE343430A1118E8987D1FE343430A1&view=detail&FORM=VIRE
Frog Fish from the Museum of New Zealand to Papa Tongarewa.

Poster with key macroinvertebrate life in the river.


My first task is to figure out to how to calibrate the Applied Biosystems QuantStudio 6.   The Applied Biosystems™ QuantStudio™ 6 Pro and 7 Pro Real-Time PCR Systems use fluorescent-based polymerase chain reaction (PCR) reagents to quantify target nucleic acid sequences.  

The instrument collects raw fluorescence data at different points during the PCR cycle, depending on the type of run performed.

The QuantStudio™ 6 Pro Real-Time PCR System has a coupled five-color filter set. 

The QuantStudio™ 6 Pro System and the QuantStudio™ 7 Pro System have interchangeable blocks. 

The calibration status applies to the combination of the instrument and the block. If a block is moved to an instrument that it has not been calibrated on, that combination of block and instrument needs to be calibrated. This applies even if the block has previously been calibrated on a different instrument. If a block is moved back to an instrument that is has previously been calibrated on, it does not need to be calibrated again. It needs to be calibrated again if the calibration is expired. A run can still be started if a calibration of the block on the instrument is expired. A run cannot be started if a block has never been calibrated on the instrument.

Calibration workflow:

Perform an ROI/uniformity calibration

You are automatically prompted to perform background calibration.

Perform a background calibration

Perform any time that ROI/uniformity calibrations are current.

Perform system dye calibrations

Perform any time that ROI/uniformity and background calibrations are current.

(Optional ) Perform custom dye calibrations Perform any time that ROI/uniformity and background calibrations are current

I will need: 
  1. ROI/Uniformity plate
  2. Background calibration plate
  3. Dye calibration plates
Calibration procedure:

Thaw, vortex, and centrifuge a calibration plate

IMPORTANT! Keep calibration plates protected from light until you perform the calibration. Do not remove the plate from its packaging until you are ready to use it. Prolonged exposure to light can diminish the fluorescence of the dyes in the wells of calibration plates.

1. Remove the calibration plate from the freezer, then thaw the plate in its packaging for 30 minutes.

IMPORTANT! Use each plate within 2 hours of thawing.

2. While wearing powder‐free gloves, remove the calibration plate from its packaging. Do not remove the optical film.

Note: Do not discard the packaging for the calibration plate. Each calibration plate can be used up to three time if the following conditions are met:

· The plate is stored in its packing sleeve at –25°C to –15°C.

· The plate is used within 6 months after opening.

· The plate is used before the plate expiry date.

3. Vortex the plate for 5 seconds, then centrifuge at 750–1,000 × g for 2 minutes.

4. Confirm that the liquid in each well is at the bottom of the well and free of

bubbles. If it is not, centrifuge the plate again.

IMPORTANT! Keep the bottom of the plate clean. Fluids and other contaminants on the bottom of the plate can contaminate the sample block and cause an abnormally high background signal.

New book arrived!

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