DNA library prep kits we offer:
- Illumina® DNA Prep, Tagmentation (previously known as Nextera DNA Flex Library Prep, Cat.Nr. 20018705), Illumina + adapters: IDT for Illumina DNA/RNA UD Indexes Set A/B/C/D Tagmentation (384 Indexes), Illumina
- Illumina® TruSeq DNA Nano library prep (Cat. Nr. 20015965), Illumina + adapters: IDT for Illumina –TruSeq DNA UD Indexes (96 Indexes) (Cat.Nr. 20022370), Illumina
-
SMARTer® ThruPLEX® DNA-Seq Kit (Cat.Nr.R400676), Takara + adapters: SMARTer® DNA Unique Dual Index Kit - 24U Set A (Cat.Nr. R400665) and SMARTer® DNA Unique Dual Index Kit - 24U Set B (Cat.Nr. R400666), Takara
| Prep type | Recommended sample amount | Min. conc. | Min. volume | Accepted buffers |
|---|---|---|---|---|
|
Illumina® DNA Prep, Tagmentation* (previously known as Nextera DNA Flex Prep) |
200 - 1000 ng |
3 ng/μl |
60 μl |
10 mM Tris (TE also OK if conc > 40 ng/μl) |
| Illumina®TruSeq Nano DNA Library Prep** |
200 ng (350 bp insert size) 400 ng (550 bp insert size) |
2 ng/μl |
110 μl |
10 mM Tris |
|
ThruPLEX® DNA-Seq Library prep*** (ChIP / low-input DNA) |
0.2 - 100 ng |
0.1 ng/μl (ChIP/degraded DNA) 1 ng/μl (gDNA) |
20 μl |
5 - 10 mM Tris
|
Input requirements for DNA library preps.
Library prep protocols
Users may choose between library preparation by transposon tagmentation (Illumina® DNA Prep, Tagmentation) or by adapter ligation based preps with fragmentation by sonication (Illumina®TruSeq Nano DNA Library Prep or SMARTer ThruPLEX® low-input).
*For Illumina DNA prep (tagmentation) submissions, samples on any one plate submitted should all fall within the following ranges of DNA amounts: 2-20 ng, 20-50 ng, 50-100 ng or 100-1000 ng (optimum) in a maximum volume of 60 µl. If sample concentrations span these ranges, samples of similar concentration should be grouped and submitted on separate plates.
**For Illumina TruSeq Nano DNA Library Prep submissions, all samples should be normalized before submitting.
***For ChIP-seq or when limiting sample amounts are available (100 pg – 50 ng), SMARTer ThruPLEX® preps should be selected.
Sample amounts
As a general rule, more DNA is better, so please provide more than the recommended sample amounts listed below if you can. Using the minimum amount increases the chances of failed preps. We may also be willing to attempt library preparation with amounts lower than the given minima, but this must be agreed with us in advance of submission, and you must nonetheless pay the cost of consumed reagents in the event of prep failure.
Concentrations must be determined by a fluorometric method such as Invitrogen´s Qubit, not spectrophotometric methods such as Nanodrop.
Samples with high genomic DNA concentrations (>1 µg/µl) cause problems with our automation due to their viscosity, so are not acceptable. Please dilute your samples to under this limit and re-check their concentration. Do not send DNA samples in less than 10 µl.
Sample integrity
Genomic DNA samples for Illumina DNA prep, tagmentation (previously known as Nextera DNA flex prep), should have high MW with no degradation. For other preps, fragmented DNA (as from FFPE samples) is acceptable, as the DNA will anyway have to undergo fragmentation by sonication prior to sample prep. Evidence of sample size & quality (gel image or bioanalyzer results) is mandatory before submission, unless agreed in advance. For genomic DNA samples, an agarose gel showing high MW DNA with no degradation is most appropriate (indicate relevant marker sizes, and amount of sample loaded). For ChIP samples, is not necessary to provide gel or Bioanalyzer documentation . However, size of input DNA prior to immunoprecipitation must be shown.
Sample purity
For DNA samples the 260/280 ratio should fall in the range 1.8-2.1 and the 260/230 ratio within 1.8-2.4. DNA for sequencing should be treated with RNase before final cleanup and submission. It is not necessary to provide 260/280 or 260/230 ratios for ChIP samples. Also note that ChIP samples must not contain salmon sperm DNA or other nucleic acid blocking agents.
Accepted buffers
See table. Generally, do not use buffers containing EDTA, as this will inhibit the enzymatic steps of sample library preparation. We do not accept lyophilized or precipitated samples. Samples delivered in inappropriate buffers will either be returned to the user or subject to additional cleanup costs of 1000 NOK/sample at our discretion.
Library preps from WGA material
Whole-genome amplified (WGA) and whole-transcriptome amplified (WTA) samples are frequently problematic, so will only be sequenced at the risk of the submitter