Reformat and Import Target Regions Into an AmpliSeq for Illumina Design
| 1. | Open the TruSeq Custom Amplicon design target regions file (*.bed) in a spreadsheet editor, such as Excel. |
| 2. | Delete the fifth and sixth columns from the file. |
| 3. | Make sure that all target names are 20 alphanumeric characters or less. Hyphens, periods, underscores, and whitespaces are allowed. |
The name field can be empty.
| 4. | Save and close the target regions file. |
| 5. | Create a new AmpliSeq for Illumina Gene design to target exons and large (> 125 bp) regions or an AmpliSeq for Illumina Hotspot design to target SNVs and small (< 125 bp) regions. See Create an AmpliSeq for Illumina Gene Design or Create an AmpliSeq for Illumina Hotspot Design. |
Make sure to select the species that matches the original TruSeq Custom Amplicon design. For human TruSeq Custom Amplicon designs, select Human (hg19).
| 6. | Configure the new AmpliSeq for Illumina design by selecting the Sample Type and Max Amplicon Length that most closely match those of the original TruSeq Custom Amplicon design. |
Use the following table for selecting equivalent amplicon sizes between the two assay types:
|
TruSeq Custom Amplicon design Amplicon Size |
AmpliSeq for Illumina custom design Max Amplicon Size |
|---|---|
|
150 |
140 |
|
175 |
175 |
|
250 |
275 |
|
425 |
375 |
| 7. | Use the default setting for Stringency. |
| 8. | Select Next. |
| 9. | Upload the TruSeq Custom Amplicon design target regions file into the AmpliSeq for Illumina design. See Add Targets by Uploading Files. |