Input Recommendations
The following section provides input recommendations.
Sample Nucleic Acid Purity
IMAP supports DNA and RNA from a variety of sample types. To improve library preparation success, use the highest quality nucleic acid input. It is good practice to characterize nucleic acid quality before beginning targeted library prep. Due to the variety and challenging nature of many sample types, it may not be possible to reach the specifications listed in the following table. If you experience difficulties in making your targeted amplicon libraries with nucleic acids that do not meet these recommendations, it may be beneficial to clean up the samples and repeat your library preparation.
|
Purity Check |
RNA |
DNA |
|---|---|---|
|
A260/280 |
2.0–2.2 |
1.8–2.0 |
|
A260/230 |
2.0–2.2 |
2.0–2.2 |
Input Amount
Given the highly variable target abundance in various sample types used with this technology, Illumina recommends adding the maximum volume of input nucleic acid permitted in this protocol. You may need to optimize the input concentration by adding solution to increase, or diluting the solution to decrease the nucleic count. When using purified RNA as the only input with this kit, Illumina recommends using a DNase treatment step during RNA purification.
Purification Kits
Many commercially available DNA and RNA extraction and purification kits are available. Choose a kit or method that produces the highest quality nucleic acids possible for your specific targets and from your specific sample types. Illumina tested the following kits, but you may find that other kits or methods work better for your samples and targets.
| • | Quick-DNA/RNA Viral MagBead, Zymo Research, part # R2141 |
| • | ZymoBIOMICS DNA/RNA Miniprep Kit, Zymo Research, part # R2002 |
| • | QIAamp Viral RNA Mini Kit, Qiagen, part # 52906 |
