Illumina NGS Libraries

We offer single paired end library construction (whole genome, library pool or clone). With our optimized low and high throughput protocols we can prepare single libraries to 96 libraries at a time. Multiple samples can be pooled to construct a single paired end library or we can construct single unique barcoded libraries from multiple samples. Libraries can be normalized and delivered separately or pooled to a singe tube after normalization.

Pools of BAC or Fosmid clones in single tube to 96-well format with high quality NGS grade DNA preparation.

This is an exciting and cost effective to inexpensively sequence many clones.

Project Flow:

Amplicon Express to construct either a Fosmid or BAC library of your organism.  We then transfect or transform and titer the ligation.  Then pool the unpicked clones in 96-well plates with ~5000 fosmid clones per well or ~1500 BAC clones per well.

We can then deliver you with 300 ng to 1.5 µg of NGS quality DNA in 96-well format.

Additionally our team can construct individually tagged 200-600 bp paired end illumina libraries normalized and ready to run on a sequencer.

If you desire to have us pick and array your library prior to pooling (or you have an existing library), this can be done as well.  We currently offer 1 to 24 pooled clones per 96-well tray formats.

We can then provide you with 300 ng to 1.5 µg of NGS quality DNA in 96-well format.  In addition we can construct individually tagged 200-600 bp paired end illumina libraries normalized and ready to run on a sequencer.

Start a Project guide for starting material requirements

Realtime PCR will accurately quantify amplifiable molecules in a NGS library. A quality check by realtime PCR amplification of NGS libraries is critical to assessing its quality in order to get proper cluster generation while sequencing in any Illumina sequencing platform. KAPA Illumina Library Quantification Kits (KAPA Biosystems) are ideally suited for this purpose.

FIG 1.

Here is a real time PCR trace for a 96-well plate illumina NGS library construction of pooled clones constructed by our NGS team.

Rows A & B = contains standards and negative controls

Rows C, D, E, F, G, & H = contains DNA libraries

Here is a real time PCR melt curve for a 96-well plate illumina NGS library construction of pooled clones constructed by our NGS team.