BioMicroCenter:Illumina Library Preparation: Difference between revisions
imported>Stuart S. Levine |
|||
| (6 intermediate revisions by 2 users not shown) | |||
| Line 1: | Line 1: | ||
{{BioMicroCenter}} | {{BioMicroCenter}} | ||
[[Image:BMC_samplePrep_pipes. | [[Image:BMC_samplePrep_pipes.jpeg|thumb|right|400px|Simplified workflow in the MIT BioMicro Center]] | ||
Generating high-quality data on | Generating high-quality data on any short read 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 short read sequencing adapters. | ||
== Short Read Library Prep Services== | == Short Read Library Prep Services== | ||
| Line 28: | Line 28: | ||
* Amplicon / 16S | * Amplicon / 16S | ||
* NexteraXT | * NexteraXT | ||
* Nextera Flex | * Nextera Flex | ||
* NEB UltraII | * NEB UltraII | ||
| Line 41: | Line 40: | ||
== OTHER SERVICES == | == OTHER SERVICES == | ||
=== SIZE SELECTION === | === SIZE SELECTION === | ||
The BMC offers [[BioMicroCenter:PippinPrep|PippinPrep Size Selection]] post library construction if custom insert sizes are needed. | The BMC offers standard SPRI cleans for NGS libraries and [[BioMicroCenter:PippinPrep|PippinPrep Size Selection]] post library construction if custom insert sizes are needed. | ||
=== QUALITY CONTROL === | === QUALITY CONTROL === | ||
All NGS libraries are quality controlled prior to use. The BioMicro Center offers two 'flavors' of quality control for | All NGS libraries are quality controlled prior to use. The BioMicro Center offers two 'flavors' of quality control for NGS libraries.<BR><BR> | ||
Standard quality control includes analysis on the [[BioMicroCenter:QC#AATI_FRAGMENT_ANALYZER|Fragment Analyzer]] and quantification by four point [[BioMicroCenter:RTPCR|qPCR]] quantification for each sample. This confirms both the sizing as well as the presence of Illumina adapters on the libraries.<BR><BR> | Standard quality control includes analysis on the [[BioMicroCenter:QC#AATI_FRAGMENT_ANALYZER|Fragment Analyzer]] and quantification by four point [[BioMicroCenter:RTPCR|qPCR]] quantification for each sample. This confirms both the sizing as well as the presence of Illumina adapters on the libraries.<BR><BR> | ||
Rapid Quality Control is less expensive and quicker. It includes analysis on the [[BioMicroCenter:QC#AATI_FRAGMENT_ANALYZER|Fragment Analyzer]] and quantification by PicoGreen on the [[BioMicroCenter:Varioskan|Varioskan]] for each sample. Final pools are quantified by [[BioMicroCenter:RTPCR|qPCR]] before loading. While this method is significantly faster and less expensive, we cannot guarantee pool balance with it. | Rapid Quality Control is less expensive and quicker. It includes analysis on the [[BioMicroCenter:QC#AATI_FRAGMENT_ANALYZER|Fragment Analyzer]] and quantification by PicoGreen on the [[BioMicroCenter:Varioskan|Varioskan]] for each sample. Final pools are quantified by [[BioMicroCenter:RTPCR|qPCR]] before loading. While this method is significantly faster and less expensive, we cannot guarantee pool balance with it. | ||
<BR><BR>[[BioMicroCenter:PREP_TEST]] | <BR><BR>[[BioMicroCenter:PREP_TEST]] | ||
Latest revision as of 17:51, 30 April 2025
Generating high-quality data on any short read 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 short read sequencing adapters.
Short Read Library Prep Services[edit]
Please follow the links in the table below for more information about our library preparation offerings.
| DNA | RNA |
|---|---|
Standard DNA Methods include
|
Standard RNA Methods include
|
High Throughput DNA Methods include
|
High Throughput RNA Methods include
|
OTHER SERVICES[edit]
SIZE SELECTION[edit]
The BMC offers standard SPRI cleans for NGS libraries and PippinPrep Size Selection post library construction if custom insert sizes are needed.
QUALITY CONTROL[edit]
All NGS libraries are quality controlled prior to use. The BioMicro Center offers two 'flavors' of quality control for NGS 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.
BioMicroCenter:PREP_TEST