0557 Extraction of 400 microliters blood on the Hamilton Chemagic star

Summary

The SOP describe the usage and maintenance of the extraction robot Chemagic STAR.

1. Purpose

To describe the usage and maintenance of the extraction robot Chemagic STAR.

2. Scope

Instruction for DNA extraction on the robots Mimmi and Musse with an automatic workflow, limited to the 400µl method.

Chemagic STAR is an extraction robot, which by using this method can extract up to 2x96 blood samples from a 2 ml deepwell plate (DWP). Mimmi consists of three parts; one extraction part with a magnetic head (Chemagen), one liquid handling part with 4x 1ml channels and 4x 5ml channels (Hamilton), and one spectrophotometer (Trinean).  Musse is the same as Mimmi, but lacks the spectrophotometer.

3. Responsibility

The person responsible for the process is also responsible for the equipment, updating this instruction, and teaching new staff how to use the robot. The staff is responsible for following this instruction while using the robot.

4. Material

  • 2 ml deepwell plate (DWP), Chemagen 6000555
  • REMP 300 container, REMP 1800503
  • Chemagic STAR DNA Blood400 Kit, Chemagen 02001765
  • Pipette tips without filters 1 ml and 5 ml, Hamilton CO-RE tips 235900, 184020
  • Waste bags for pipette tips, Hamilton 199202

5. Procedure extraction

5.1 Turn on the extraction and liquid handling parts of the robot by pressing the two switches: one on the front and the other one behind the door. Also, turn on the computer if has been shut down.

5.2 Check that the Tip Eject Plate (the metal plate where the tips get ejected) and the waste bag are at place and in the right position, i.e. right below the holder.  

5.3 Load the working table with all the necessary material, as in the picture below. Make sure all pipette tips holders are stuck in the rack and that all plates are standing straight. 

To load the sleeves and the elution plates in the back, the ”load/unload” script can be used:

  • Place the sleeves and elution plates as marked on the working table.
  • Click ”loading unloading plates” on the desktop.
  • Choose ”load” or ”unload” and click ”ok”.

5.4 Important to know about the different solutions used:

  • Solutions nr. 2-5 are stable in the tray up to 4 hours. Longer storage without lid is not recommended, as the ethanol evaporates.
  • The protease in its dissolved form is stable at 4oC up to 4 weeks. For longer storage, store small aliquots at -20oC (aliquots that have been thawed should not be frozen again).
  • The magnetic beads must be resuspended before each use by shaking the bottle heavily.

    For more information on the included buffers in this kit, see ”Chemagic STAR DNA Blood 400 kit

5.5 Start the run by clicking ”Extraction”.

5.6 Tick the box ”Start application” and ”reset tipcounter” if you have refilled the tips.  Click ”ok”.

5.7 Wait until the following box appears and choose ”1 plate” or ”2 plates” depending on how many samples you want to run (96 or 192 samples).
 

5.8 When the following box appears, click “yes”.


 

5.9 If you have refilled the tips, answer the questions whether there are enough tips or not.

 

5.10 The robot will now scan the plates and the containers. The LIMS will validate the information from the scanning and tell which wells should be extracted and not, or start immediately if all wells are ok.

5.11 To start the extraction click ”ok”.

5.12 When the extraction is finished, the following box will show up.

5.13 Check that all samples have been transferred to the REMP-container, and that the samples are in the LIMS, ses LIMS user manual.

5.14 The concentration of the samples then needs to be measured, see the instruction “Concentration measurement Trinean” (”Koncentrationsmätning Trinean”).

5.15 The plates with blood and sleeves should be thrown in the risk waste. The other plates are emptied and then thrown in the normal waste. The sleeve plate is recycled as plastics.

6. Maintenance

Daily maintenance

6.1 Clean the head of the channels with 70 % ethanol by wetting a paper towel and then dry clean the metal part at the bottom of the channel. Be careful with the o-ring above, as it dries out from ethanol. Clean the Tip Eject Plate with water and detergent.

6.2 Throw away what is left of the reagents and wash the tray by rinsing it in water, finishing with deionized water.

Weekly maintenance

6.3 When needed, clean the racks with water and detergent.

6.4 Wipe off the working table with water and some detergent. If needed disinfectant can be used, but make sure the table is clean first, otherwise you just smear the dirt.

6.5 Wipe off the outer working table with disinfectant. It can also be lifted and rinsed with water.

6.6 Wipe off the Auto-load band and the Auto-load with a paper towel wetted with water.

6.7 Clean the 2D Barcode Scanner with lint-free paper towels and ethanol when needed.

6.8 Note the maintenance in the logbook.

7. Errors

7.1 If the 2D Barcode Scanner cannot read all 2D barcodes the following message will appear. Change the tube and click ”ok”.

7.2 If the extraction is interrupted all the sleeves must be removed from the magnetic head.
Click on ”MSN.exe” on the desktop. Log in as Hamilton, password Hamilton, and choose the protocol ”remove 96 tips in pos.1.che” and click ”start”.

 

Citation

SOP collection, KI Biobank. 0557 Extraction of 400 microliters blood on the Hamilton Chemagic star. The Molecular Methods Database. 2016/01/25

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