BioMicroCenter:PREP TEST

From OpenWetWare
Jump to navigationJump to search

HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY

Library Preparation Services

Sample Preparation available in BioMicro

Generating high-quality data on the Illumina sequencing platform requires high-quality libraries. The BMC currently offers library preparation services for a variety of starting materials. Prior to sequencing, all samples must pass the BioMicro Center’s Sequencing Quality Control process, which verifies selection of inserts of a desired size and correct ligation of Illumina adapters.

Illumina Library Prep Services

Please follow the links in the table below for more information about our library preparation offerings.

METHOD TYPE DNA RNA
STANDARD Ligation-mediated PCR Methods

Tagmentation Methods

Standard Input

Low Input

Small RNA upon request

HIGH THROUGHPUT Ligation-mediated PCR Methods
  • NEB Ultra II

Tagmentation Methods

  • Nextera Flex
  • Nextera XT

Amplicon Methods

  • 16S Metagenomic
  • Custom Amplicon
Standard Input
  • NEB Ultra II Directional RNA with:
       Poly(A) mRNA Isolation
       rRNA Depletion (H/M/R)
  • 3' Digital Gene Expression (3'DGE)

Low Input

  • Clontech SMART-Seq v2
       - Poly(A) based selection
  • Clontech ZapR
       - rRNA depletion based selection

OTHER SERVICES

SIZE SELECTION

The BMC offers PippinPrep Size Selection post library construction if custom insert sizes are needed.

QUALITY CONTROL

All NGS libraries are quality controlled prior to use. The BioMicro Center offers two 'flavors' of quality control for Illumina libraries.

Standard quality control includes analysis on the Fragment Analyzer and quantification by four point qPCR quantification for each sample. This confirms both the sizing as well as the presence of Illumina adapters on the libraries.

Rapid Quality Control is less expensive and quicker. It includes analysis on the Fragment Analyzer and quantification by PicoGreen on the Varioskan for each sample. Final pools are quantified by qPCR before loading. While this method is significantly faster and less expensive, we cannot guarantee pool balance with it.