0899 Trinean Concentration measurement

Summary

The document describe how to use the Trinean spectrophotometer, placed together with the Hamilton Chemagic STAR extraction robot (Mimmi) and the robot Hamilton Microlab Star LET (Pärlan) at KI Biobank.

1.   Purpose

To describe the usage of the Trinean spectrophotometer, placed together with the Hamilton Chemagic STAR extraction robot (Mimmi) and the robot Hamilton Microlab Star LET (Pärlan).

2.   Scope

Instruction for DNA quantification with the Trinean DropSense-96 in the automated process for large-scale extraction.

3.   Responsibility

  • The process responsible person is responsible for the equipment together with the medical technical engineer.
  • The process responsible person is responsible for updating this instruction and teaching new personnel how to use the robot. 
  • The personnel is responsible for following this instruction while using the robot.

4.   Material

  • Trinean DropPlate96-S, Techtum 27-216096-S
  • Pipette tips 10µl without filter, Hamilton CO-RE tips 235900
  • Pipette tips 50µl without filter, Hamilton CO-RE tips 235966
  • Trash bags for pipette tips, Hamilton 199202
  • Pipette 10µl, Finntip
  • Pipette tips 10µl, Finntip 1144-8319

5.   Measurement on Pärlan

  1. Two different methods can be run on Pärlan. Method 1 is used for REMP-300 and Method 2 is used for REMP-900. Choose the method on the desktop on the computer (different icons).
  2. Method 1 (REMP-300): Place the REMP-300 containers to be measured in the right position on the working table. Also place the number of DropPlates and pipette tips needed on the working table (see picture below). Note! It is very important that the DropPlate is placed with the label outwards, i.e. the position A1 up in the left corner, otherwise the plate will break in the spectrophotometer.  
  3. Turn on the liquid handling part of the robot using the green button on the left side and the spectrophotometer on the backside. Turn on the computer if has been shut down.
  4. Click on ”Trinean DropQuant” on the desktop.
  5. Log in as ”remote” without password.
  6. When this picture appears, click on ”Concentration” on the desktop.
  7. Click on the green arrow in the software or F5 to start the concentration measurement.
  8. Choose how many plates to be measured, then click ok.
  9. When the following box appears: Choose in which position the tubes are, then click ok.
  10. When the following box appears – make sure the pipette tips have been marked by marking the position in the picture. Alternatively, tick/untick the “Remove all”. Make sure there are tips left under “Remaining”, and that they are of the right type.
  11. The robot will now scan the plate/s. When the LIMS has validated which samples to measure, the pipetting will start automatically. The method is finished when ”End method- completed” is displayed on the last row in the ”trace wiew” window.
  12. Method 2 (REMP-900): Choose which type of plate you are using; REMP-900. Note! 5.3 – 5.7 is the same as in method 1.
  13. Place the REMP-900 containers to be measured in the right position on the working table. Also place the number of DropPlates and pipette tips needed on the working table (see picture below). Note! It is very important that the DropPlate is placed with the label outwards, i.e. the position A1 up in the left corner, otherwise the plate will break in the spectrophotometer.  
  14. When the following box appears: Choose in which position the tubes are, then click ok.
  15. When the following box appears – make sure the pipette tips have been marked by marking the position in the picture. Alternatively, tick/untick the “Remove all”. Make sure there are tips left under “Remaining”, and that they are of the right type.
  16. The robot will now scan the plate/s. When the LIMS has validated which samples to measure, the pipetting will start automatically. The method is finished when ”End method- completed” is displayed on the last row in the ”trace wiew” window.
  17. Approve the run in the LIMS, see LIMS user manual (LIMS användarmanual).
    To approve a batch, there should be results for all samples in the LIMS and only a limited number of samples should have a concentration <50 ng/µl and a ratio outside the interval 1.7-2.3.
  18. The samples that need to be measured again, samples with no concentration measured, are stored in another REMP-container in the fridge with the instrument id 013 in room 1427, see LIMS user manual (LIMS användarmanual). When the container is full, the samples are measured, and a new container is filled with samples to be measured again.

6.   Measurements on Mimmi

  1. Place the REMP containers to be measured in the right position on the working table. Also place the number of DropPlates and pipette tips needed on the working table (see picture below). Note! It is very important that the DropPlate is placed with the label outwards, i.e. the position A1 up in the left corner, otherwise the plate will break in the spectrophotometer
  2. Turn on the liquid handling part of the robot using the green button on the left side and the spectrophotometer on the backside. Turn on the computer if has been shut down.
  3. Click on ”Trinean DropQuant” on the desktop.
  4. Log in as ”remote” without password.
  5. When this picture shows up, click on ”Concentration” on the desktop.
  6. Click on the green arrow or F5 to start the concentration measurement.
  7. Choose an alternative: 0/1. Press: OK.
  8. Choose ”Low Volume (96 samples)”, click ”ok”.
  9. When the following box appears:

    Choose how many plates to measure, click ok.
  10. When the following box appears:

    Choose in which position the tubes are, then click ok.
  11. The robot will now scan the plate/s. When the LIMS has validated which samples to measure, the pipetting will start automatically. The method is finished when ”End method- completed” is displayed on the last row in the ”trace wiew” window.
  12. Approve the run in the LIMS, see LIMS user manual (LIMS användarmanual).
    To approve a batch, there should be results for all samples in the LIMS and only a limited number of samples should have a concentration <50 ng/µl and a ratio outside the interval 1.7-2.3.
    The samples that need to be measured again, samples with no concentration measured, are stored in another REMP-container in the fridge with the instrument id 013 in room 1427, see LIMS user manual (LIMS användarmanual). When the container is full, the samples are measured, and a new container is filled with samples to be measured again.

7.   Quality control

  1. Our internal quality control QC-DNA, see Quality controls for DNA quantification (Kvalitetskontroller för DNA kvantifiering) is measured once per week. This measurement can only be done manually.
  2. Pipette 8 blank samples (eluation buffer, Chemagen extraction kit) and 8 QC-DNA samples on a DropPlate96-S, 3µl in each well.
  3. Click on DropQuant on the desktop and log in as lab manager with the same password.
  4. Click on ”New Measurement”.
  5. Click ”next”.
  6. Click in the picture of the plate on the screen and mark which samples are blanks and which ones are QC-DNA. To change sample type, click in the ”Blank” field.
  7. Click ”Start Test”.
  8. The spectrophotometer will now open and the sleigh come out. Place the DropPlate with the label against the wall, i.e. position A1 in the lower right corner. Note that it is important to position the plate correctly, otherwise it might break during the measurement.  
  9. When the measurement is finished, choose “export file to excel” (”exportera fil till Excel”). Save the measurement under : P:\Biobank\SSB 2010\Utrustning\Hamilton\QC-DNA.

8.   Maintenance

No maintenance is needed on the spectrophotometer. For the liquid handling part of the robot, see “Instruction – Maintenance Hamilton, 0832-2) (”Instruktion – Underhåll Hamilton, 0832-2) in Centuri.

9.   Errors

Error 17- The sample has not been pipetted and needs to be measured again.  

Error 102 - Problem during pipetting; approve if the result is within the correct interval, i.e. above 50 ng/µl and the ratio between 1.7-2.3. If not, the measurement is run again.

 

 

Citation

SOP collection, KI Biobank. 0899 Trinean Concentration measurement. The Molecular Methods Database. 2016/01/26

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