Exporting to the EXQuest spot cutter
REDFIN is fully integrated with the EXQuest spot cutter from Bio-Rad and this manual will show you step-by-step how to export your data from REDFIN for use in EXQuest.
Step 1: Filter your list
Before proceeding to the exporting wizard you need to ensure that your list of spots only contains those you wish to export for identification.
If you have previously star marked those spots you can now simply filter on the star rating. Otherwise, use an appropriate combination of filters until only those spots you wish to export remain in the list.
Step 2: Click on the "export to picker" icon
You are now ready to export your data to the EXQuest spot cutter.
Click the little pipette icon in the top right corner of your screen.
Choose EXQuest from the drop down menu.
Step 3: Select to cut from a new gel
As you are cutting the spots from a new, preparative gel, select the "A new gel" radio button.
Click Start.
Step 4: Upload your preparative gel
You now need to upload your new, preparative gel into the REDFIN software and align it to one of the analyzed gels in the project. This will allow the software to triangulate the picking coordinates to the new gel for precise excision.
Start by clicking the Load gel button. A window will pop up allowing you to navigate to the location of your preparative gel on your computer. Select the gel and upload it to REDFIN.
Next you need to choose one of your analyzed gels as a reference gel for alignment.
Click on the drop-down menu next to "Select existing gel:" and choose a gel from the list. Make sure to select the gel that is most similar to your new gel.
Click Next to proceed to the alignment step.
Step 5: Align your new gel
REDFIN has now transferred the spot centers of the spots you wish to cut (indicated by red dots) from the analyzed gel to the new gel. However, as the new gel has not been aligned to the analyzed gel, the spot markers are probably slightly off. To correct for this, you need to align the new gel to the analyzed gel.
First, check the include missing spots check box to show also spots that have not been detected in your chosen analyzed gel. A red square indicates missing spots.
Next, make sure you have sufficiently visualized the spot pattern by adjusting the contrast and brightness controls underneath the image.
Now click on a red spot center in the analyzed gel (right). A green cross will appear indicating an alignment marker.
Next, click on the corresponding spot in the new gel to place an alignment marker there. Note, that you need to click on the center of the corresponding spot, not the red dot! As soon as you have placed the alignment marker, the image will align and the red spot marker should now be on the center of the spot.
To delete an alignment marker (green cross or green circle) simply right click on it.
You can zoom in and out of the gel by either clicking on the "-"and "+" magnifying glasses or by scrolling your mouse wheel.
You can move around the zoomed image by switching to the hand tool. You do this by either clicking on the hand symbol or by holding down the space bar.
To return to alignment mode click again on the hand symbol or release the space bar.
Continue aligning the gels until all red spot markers are on the centers of the spots you wish to cut from the gel. You may focus only on the gel areas containing spot markers. It is not necessary to align the entire gel.
Click Next to proceed to exporting.
Step 6: Review and export
You are now presented with the new gel that you will use for spot excision. Check that all red spot markers are indeed on the center of the spots you wish to cut.
If you see anything that needs correcting, click Previous to return to the alignment step.
Once you have verified that everything is correct, click the Export button.
A pop-up window will appear allowing you to navigate to the location on your computer where you wish to save your EXQuest spot cutter file.
Save your spot cutter file.
