BioMicroCenter:Illumina Library Preparation: Difference between revisions
No edit summary |
No edit summary |
||
| (7 intermediate revisions by 3 users not shown) | |||
| Line 1: | Line 1: | ||
{{BioMicroCenter}} | {{BioMicroCenter}} | ||
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 libraries 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. | |||
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 | |||
== Short Read Library Prep Services== | == Short Read Library Prep Services== | ||
| Line 8: | Line 7: | ||
{| style="wikitable" border=1 | {| style="wikitable" border=1 | ||
! DNA | !width=300| DNA | ||
! RNA | !width=300| RNA | ||
|- | |- | ||
| '''[[BioMicroCenter:DNA_LIB|Standard DNA Methods include]]''' | | '''[[BioMicroCenter:DNA_LIB|Standard DNA Methods include]]''' | ||
| Line 20: | Line 19: | ||
| '''[[BioMicroCenter:RNA_LIB|Standard RNA Methods include]]''' | | '''[[BioMicroCenter:RNA_LIB|Standard RNA Methods include]]''' | ||
* NEB UltraII <br> polyA stranded method | * NEB UltraII <br> polyA stranded method | ||
* NEB ribosomal depletion | * NEB ribosomal depletion <br> stranded rRNA depletion | ||
* | * Clontech SMARTseq <br> Low input polyA based | ||
* Clontech ZapR <br> Low input rRNA depletion based | * Clontech ZapR <br> Low input rRNA depletion based | ||
* | * Small RNA prep | ||
|- | |- | ||
| '''[[BioMicroCenter:DNA_HTL|High Throughput DNA Methods include]]''' | | '''[[BioMicroCenter:DNA_HTL|High Throughput DNA Methods include]]''' | ||
* Amplicon / 16S | * Amplicon / 16S | ||
* NexteraXT | * NexteraXT | ||
* Nextera Flex | * Nextera Flex | ||
* NEB UltraII | * NEB UltraII | ||
| Line 37: | Line 35: | ||
* SMARTseq v2 | * SMARTseq v2 | ||
* ZapR | * ZapR | ||
|} | |||
== Long Read Library Prep Services == | |||
The BioMicro Center also supports library preparation for long-read sequencing on two platforms. See the [[BioMicroCenter:LongRead|Long Read Sequencing page]] for full details on sequencing services and platform comparison. | |||
{| style="wikitable" border=1 | |||
!width=300| Oxford Nanopore Technologies | |||
!width=300| PacBio | |||
|- | |||
| '''[[BioMicroCenter:LongRead#PromethION_Library_Preparation|Nanopore Library Methods include]]''' | |||
* LSK114 — Ligation-based dsDNA | |||
* RBK114 — Tagmentation-based / ultra-long | |||
* RNA004 — Direct RNA sequencing | |||
* Native barcoding available (up to 96 samples) | |||
| '''[[BioMicroCenter:LongRead#Revio_.E2.80.94_Library_Preparation|PacBio Library Methods include]]''' | |||
* SMRTbell Prep Kit v3.0 | |||
* Genomic DNA (HiFi, 10–20 kb reads) | |||
* Size selection via PippinPrep available | |||
|} | |} | ||
== 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 22:55, 23 April 2026
HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY
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 libraries 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
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
|
Long Read Library Prep Services
The BioMicro Center also supports library preparation for long-read sequencing on two platforms. See the Long Read Sequencing page for full details on sequencing services and platform comparison.
| Oxford Nanopore Technologies | PacBio |
|---|---|
Nanopore Library Methods include
|
PacBio Library Methods include
|
OTHER SERVICES
SIZE SELECTION
The BMC offers standard SPRI cleans for NGS libraries and 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 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