Library pooling
Indexing
To pool multiple libraries in the same sequencing run, each library needs to have a unique index (barcode). Barcoding strategies can vary by application and library type. Standard Illumina sequencing services will ‘demultiplex’ the dataset and distribute fastq file(s) based on the Illumina Index reads that capture library barcode information.
Pooling
If you submit barcoded libraries as separate samples, we can pool libraries prior to sequencing.
- Simple pooling for high throughput projects (libraries in 96 well plate format): equal volumes of each library are pooled, and may not generate equal reads per sample during Illumina sequencing. Custom pooling is designed to more carefully balance reads within a sequencing run, and requires quantification and size QC (or dPCR) to estimate molarity prior to pooling.
- Custom pooling is based on an estimate of library molarity to balance the reads per sample during sequencing. Library molarity can be estimated from quantification and size distribution, or using dPCR for higher accuracy (all incur additional charges). A typical goal is to pool equimolar amounts of each library to generate approximately equal numbers of reads per sample. Libraries can also be pooled with custom balancing targets, and/or pooled in anticipation of a Pippin size selection on the final pool. Custom pooling will be charged based on time required (custom lab work), or per 96-well plate.
Pooling may require a consultation to ensure the appropriate approach is used; contact us for more information.
Service pricing
Description | Internal price (Cornell and Cornell affiliates) | External price |
---|---|---|
Simple pool (per 96 well plate, constant vol per well) | $55 | $90 |
Custom pool (per 96 well plate, custom vol per well) | $90 | $150 |
Custom lab work (per hour) | $55 | $90 |
Description | Internal price (Cornell and Cornell affiliates) | External price |
---|---|---|
Digital PCR: Illumina library quantification - per sample | $30 | $50 |
Fragment Analyzer (RNA or DNA analysis) - per sample | $15 | $25 |
Pippin size selection (first sample) | $30 | $50 |
Pippin size selection (samples 2-12) | $25 | $40 |
DNA cleanup (96 well plate) | $275 | $450 |
DNA cleanup (per sample) | $25 | $40 |
Nucleic acid quantification (96-well plate) | $135 | $220 |
Nucleic acid quantification (per sample) | $5 | $8 |