Tapestation Protocol

Tapestation Protocol

Available Tapestation Assays

  • RNA
  • High Sensitivity RNA
  • Genomic DNA
  • D1000 DNA
  • High Sensitivity D1000 DNA
  • D5000 DNA
  • High Sensitivity D5000 DNA

Preparing Samples

Retrieve the screentape, sample buffer, and ladder (if necessary) for your chosen assay at least 30 minutes before starting
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All reagents are in the Beiting Lab 4°C fridge, on the door
If you are doing an RNA or HS RNA assay, set the thermocycler on the the “72C 3” protocol (72°C hold, 72° for 3 minutes, 12° hold)
Retrieve Tapestation strip tubes and caps from the drawer labeled “Tapestation Supplies”
Label Tapestation strip tubes on the side of the tube (do not forget to include a ladder if required)
Add the correct volume of sample buffer and sample to each tube

Sample Loading

Assay
Sample Buffer (µL)
Sample (µL)
Ladder (µL)
RNA
5
1
NA
HS RNA
1
2
NA
Genomic DNA
10
1
1
D1000
3
1
NA
HS D1000
2
2
NA
D5000
10
1
1
HS D5000
2
2
2
Cap all the tubes tightly and vortex on the IKA MS3 Vortex
Centrifuge briefly

For RNA and HS RNA Assays Only

Place tubes in the thermocycler (holding at 72°C)
Press “SKIP” to go to the next step (72°C for 3 minutes)
After the samples reach 12°C, remove the tubes from the thermocycler and place on ice for 2 minutes
After the 2 minutes on ice, centrifuge briefly again

Loading the Tapestation

Make sure the Tapestation and computer are both on and that they are connected to each other
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Login for the computer is written on the computer
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The program for the Tapestation is called “Tapestation Controller” on the computer Desktop
Very carefully (without disrupting the liquid), remove the caps from the tubes and load the tubes (in the correct orientation) in the tube location
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If you are doing a plate, load the plate in the plate location
Load the screentape in the slot with the flashing light (load the screen tape with the colored side of the tape facing you and the star burst/dot symbol on the top)
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Screentapes can be reused and are fine to run past their expiration date. Please just make sure that you have enough lanes on the tape you are using.
Ensure that there are enough pipette tips in the machine (there are more pipette tips in the “Tapestation Supplies” drawer)
Close the lid of the Tapestation
On the computer, mark off the correct number of tubes you will be running (remember to include the ladder if you are using one)
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If you are not using a ladder and position A1 indicates a ladder, you can turn it off by right clicking once on position A1 and selecting “Use Electronic Ladder”
Give each of your samples an ID or name
Press “Start”
Press “Proceed”

Saving Your Results

The “Tapestation Analysis Software” program will open automatically when the run finishes
Once you have seen your results, click on the “File” tab at the top
Click “Create Report”
Click “Save As”
Save as a PDF in the “Tapestation Data” folder
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The file already has the date and time of the run. Please add your name and some identifiers for your run, separated by underscores.