Creating the PCR Plate

The extracted DNA is transferred to the PCR plate in exactly the same arrangement, i.e. the samples remain in the same wells. For creating a PCR plate, use the search function and find the extraction plate and mark it:

Click again on New Reaction in the navigation bar. This time set Type of reaction = PCR in the pop-up window and make sure that Create plate from existing documents is selected so that the sample IDs are taken over from the extraction plate.

A view of the PCR plate opens: first of all enter the plate name in the format CaBOL_PCR_xxxx. The plate number is the same as that of the extraction plate.

Now the Lab information can be stored like we did before for the extraction plate: Select the used PCR program under Thermocycle.

Click on Edit All Wells to open a new window:

Select Locus and enter the Date, the Primer used and the Technician. The primer sequences are available in the Shared database in the folder Primer.

Press OK to close the Edit Wells window and return to the plate view .

Click on Attach GEL Image to open a new window where a gel image can be inserted.

Click on Add to select the corresponding gel image from the storing location.

Via Split GEL the plate positions are mapped onto the gel image.

Enter the number of rows and columns and the starting position and draw a frame around the place on the gel image that belongs to each well. In this example, it is done row by row. Use the Toggle Direction button if the well number in the frame is not according to the well of the plate. Each time, press OK to return to the previous Edit Gel Images window.

Based on the gel images, the Reaction State of the samples is automatically marked as passed (green) or failed (red). This classification can – and often should – be corrected manually by double clicking on the well because in some cases the automatic classification by the program seems to be a bit incomprehensible.

Based on my experience in this example I would definitely classify A1, B9, and E3 promising gel signals (passed). There is also the option to set reactions on the state suspect what I would do for D12 in this example. Select Reaction State = run for the negative control.

Close the plate view with OK to save the new plate .