BioMicroCenter:DNA HTL

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SEQUENCING --

LIBRARY PREP --

HIGH-THROUGHPUT --

COMPUTING --

OTHER TECHNOLOGY

The BioMicro Center offers high-throuhgput DNA library preparation for NexteraXT samples and for metagenomics/amplicon generation. High-throughput library generation is different from standard library preparation in a couple key ways. First, with dozens of samples to prepare, small numbers of samples may fail and the cost of reprepping those samples is NOT included in the quoted cost. For some experiments, repreps can be done by hand, but at a higher rate. Second, we are focused far more on reducing price than for standard library preparation, so some "routine services" - such as quality control or arraying of samples - is not built in.

High-Throughput Nextera XT

To minimize library prep costs for standard Illumina libraries, we have automated Illumina NexteraXT on our Tecan EVO150s. Libraries will be built to include variable i5 and i7 indexes based on the grid so dual indexing is required during sequencing of these libraries. For more information about how Nextera works, please check out the standard Nextera library preparation section of standard DNA library prep.

Plate setup
Batch Size	16
Sample Layout	Columns from left
Volume	>7ul
Concentration	0.2ng/ul*
Buffer	H2O or 10mM Tris 8.0. no organics!
Plate	Axygen 96well (CAT#)
Additional services available can be added if samples are not submitted as above	Sample setup (per 48) SPRI cleanup (if not clean in proper buffer) Advanced Analytical (if not quantified).
* For short PCR fragments, lower concentrations are required. Please speak with BMC staff for amplicons.

16S / Amplicon Sequencing

Plate setup
Batch Size	48
Sample Layout	Columns from left
Volume	>5ul
Concentration	Iniital qPCR for target has Ct < 20
Buffer	H2O or 10mM Tris 8.0. no organics!
Plate	Axygen 96well (CAT#)
Additional services available can be added if samples are not submitted as above	Sample setup (per 48) SPRI cleanup (if not clean in proper buffer)