BioMicroCenter:Illumina Library Preparation: Difference between revisions
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[[Image:BMC_samplePrep_pipes.png|thumb|right|400px|Sample Preparation available in BioMicro]] | [[Image:BMC_samplePrep_pipes.png|thumb|right|400px|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 | 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. | ||
== | |||
== Short Read Library Prep Services== | |||
Please follow the links in the table below for more information about our library preparation offerings. | Please follow the links in the table below for more information about our library preparation offerings. | ||
{| border=1 | {| style="wikitable" border=1 | ||
! DNA | ! DNA | ||
! RNA | ! RNA | ||
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| '''[[BioMicroCenter:DNA_LIB|Standard DNA Methods include]]''' | | '''[[BioMicroCenter:DNA_LIB|Standard DNA Methods include]]''' | ||
* LMPCR methods | * LMPCR methods | ||
** | ** NEB UltraII | ||
* Tagmentation Methods | * Tagmentation Methods | ||
** | ** Nextera XT | ||
** Nextera Flex | |||
| '''[[BioMicroCenter:RNA_LIB|Standard RNA Methods include]]''' | | '''[[BioMicroCenter:RNA_LIB|Standard RNA Methods include]]''' | ||
* | * NEB UltraII <br> polyA stranded method | ||
* | * NEB ribosomal depletion (NEB or Lexogen for Bacteria) <br> stranded rRNA depletion | ||
* | * SMARTer_Low-Input <br> Low input polyA based | ||
* | * Clontech ZapR <br> Low input rRNA depletion based. | ||
* | * smallRNA prep | ||
|- | |- | ||
| '''[[BioMicroCenter:DNA_HTL|High | | '''[[BioMicroCenter:DNA_HTL|High Throughput DNA Methods include]]''' | ||
* Amplicon / 16S | * Amplicon / 16S | ||
* NexteraXT | * NexteraXT | ||
* low input NexteraXT | * low input NexteraXT | ||
| '''[[BioMicroCenter:RNA_HTL|High | * Nextera Flex | ||
* NEB UltraII | |||
| '''[[BioMicroCenter:RNA_HTL|High Throughput RNA Methods include]]''' | |||
* PolyA (NEB UltraII) | |||
* NEB ribosomal depletion | |||
* High Throughput 3' Digital Gene Expression (HT3DGE) | * High Throughput 3' Digital Gene Expression (HT3DGE) | ||
* SMARTseq v2 | |||
* ZapR | |||
|} | |} | ||
== 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 [[BioMicroCenter:PippinPrep|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.<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. | |||
<BR><BR>[[BioMicroCenter:PREP_TEST]] | |||
[[ | |||
Latest revision as of 09:15, 30 March 2023
HOME -- | SEQUENCING -- | LIBRARY PREP -- | HIGH-THROUGHPUT -- | COMPUTING -- | OTHER TECHNOLOGY |
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.
Short Read Library Prep Services
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
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.
BioMicroCenter:PREP_TEST