DNA library preps

 

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. Concentrations must be determined by a fluorometric method such as Invitrogen´s Qubit, not spectrophotometric methods such as Nanodrop. It is not recommended to submit less than 100 ng DNA except in the case of small (e.g. bacterial) genomes. If your samples do not meet the requirements above, contact us and we will try to find a solution.

DNA for sequencing should be treated with RNase before final cleanup and submission.

 

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.  

 

ChIP DNA library preps

It is not necessary to provide gel or Bioanalyzer documentation of ChIP sample size or quality, nor 260/280 or 260/230 ratios. However, it is necessary to document the size of input material used prior to immunoprecipitation.

Note that ChIP samples must not contain salmon sperm DNA or other nucleic acid blocking agents.

 

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.

Published Mar. 20, 2020 11:09 AM - Last modified Feb. 23, 2022 10:19 AM