DNA-Seq methods differ in terms of the DNA fragmentation method, genome size of interest, use of PCR amplification in library creation, and the use of “mate-pair” technology. All of the protocols listed in this section utilize Illumina reagents and protocols.
This is a “standard” NGS workflow: 1) acoustic shearing of DNA, 2) enzymatic repair/ligation of adaptors, and 3) PCR amplification.
This “transposase” method does not involve physical fragmentation of gDNA, but instead uses direct incorporation of Illumina adaptors via “tagmentation”.
This protocol employs tagmentation and a first round of specialized Nextera-based intra-molecular circularization, followed by shearing and traditional TruSeq library creation, to generate “jumping libraries” of fragments that originate from a known distance apart in the original gDNA.
This workflow is tailored to small genomes, such as those of bacteria. In both Nextera and Nextera XT methods, PCR is used to amplify successful tagmented molecules.
This method is designed to avoid GC-bias associated with methods such as TruSeq and Nextera.
Contact Elyse Froehling: email@example.com.
|TruSeq Nano DNA Library Creation.|
|Input DNA requirements: 100 ng for insert size||1-any||sample||$91.12||$105.16|
|of 350 bp, 200 ng for insert size of 550 bp.|
|Nextera Library Creation.|
|Standard Scale. 50 ng of input DNA needed.||1-24||sample||$133.08||$153.29|
|High Scale. 50 ng of input DNA needed.||25-any||sample||$122.42||$141.00|
|Nextera Mate-pair Library Creation.|
|Gel Free. Standard Scale.¹||1-any||sample||$292.09||$337.69|
|Gel Plus. Standard Scale.¹||1-any||sample||$644.27||$743.02|
|Nextera XT Library Creation.|
|Input DNA requirements: 1 ng||1-24||sample||$78.62||$90.47|
|PCR-Free Library Creation.|
|1 ug of input DNA for a library insert size||1-any||sample||$132.79||$152.95|
|of 350 bp. 2 ug of input DNA for a library|
|insert size of 550 bp.|
|1. There are two different approaches to Nextera Mate-pair Library Creation: Gel-Free vs Gel-Plus. The Gel-Free protocol, which does not include fragment sizing, requires 1 μg of DNA, is more robust, and yields a higher diversity of fragments with a broader range of fragment sizes. The Gel-Plus protocol, which includes gel-based fragment sizing, requires 4 μg of DNA, and allows the selection of a narrower range of fragment sizes.|
Please contact firstname.lastname@example.org for project specifications.
Once project details are finalized, complete appropriate submission form for submitting samples or submitting client-made libraries and email to email@example.com.
1-202 Nils Hasselmo Hall (Minneapolis campus)
1-210 Cancer & Cardiovascular Research Building (Minneapolis campus)
20 Snyder Hall (St. Paul campus)
Samples should be frozen and shipped on dry ice in a 96-well plate. We recommend using plate tape to seal the wells. Place the plate inside of a plastic bag prior to placing on dry ice. Please give advance notice of submission date and time so staff can be prepared to receive samples. If shipping samples from outside the University of Minnesota, ship via express shipping carrier on dry ice to the address below.
Please send the tracking information to firstname.lastname@example.org.
University of Minnesota Genomics Center
1475 Gortner Avenue
28 Snyder Hall
St. Paul, MN 55108
Generally, turnaround is 6-8 weeks for standard NGS projects. However, please contact email@example.com for specific project turnaround time.