Sample Requirements
NGS Samples for Library Preparation
Submit samples in 1.5 ml Lo-bind microfuge tubes, labeled clearly with order number, submission date, and sample identity. Sample quantification is required prior to submission.
NOTE: Concentrations determined by NanoDrop are very unreliable. Please quantify the DNA and RNA concentrations using fluorometry. If you cannot do fluorometry, please send us 2X the minimum amount that NanoDrop determines.
| Procedure | Minimum Quantity | RNA Integrity (RIN) Score | Concentration |
|---|---|---|---|
| RNA-Seq Samples mRNA Sequencing RNA should be treated with DNAse prior to quantification |
5 μg | > 8.0 | ~100 ng/μl |
| Whole Transcriptome RNA should be treated with DNAse prior to quantification |
2 μg | > 8.0 | ~100 ng/μl |
| Illumina Small RNA Seq | 2 μg | > 8.0 | ~200 ng/μl |
| ChIP-Seq Samples DNA should be sheared to 200-600 bp |
5 ng for the ChIP DNA Sample 50 ng for the input DNA Sample DNA Integrity: Provide gel image of your input DNA with size standards |
||
| Exome Sequencing | 1-5 μg DNA Integrity: Provide gel image of your input DNA with size standards |
~100 ng/μl | |
| Whole Genome Sequencing | 2 μg DNA Integrity: Provide gel image of your input DNA with size standards |
~100 ng/μl |
Prepared Libraries
Before making your libraries, please make sure your barcodes are compatible with Illumina Standards (for details, see page 12 of Illumina's TruSeq® Sample Preparation Pooling Guide). We discourage the use of investigator-designed barcodes.
Minimum concentration: 2 nM (measured by fluorometry)
Minimum volume: 15 μl
If your samples do not meet these requirements, you will need to pool your samples into lane pools at 2 nM. We will need 20 μl of this pool.
Unfortunately we cannot offer any guaranteed cluster yield or quality for libraries prepared outside of the McDermott Center NGS Core, but we are happy to answer any questions you have regarding library preparation.