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Invitrogen™ Collibri™ Stranded RNA Library Prep Kit for Illumina™ Systems

Catalog No. A39005096
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No. of Reactions:
24 Reactions
96 Reactions
Includes:
RNA Prep and UD Index
RNA Prep Kit Only
RNA Prep, Depletion Kit
RNA Prep, Depletion, UD Index
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Catalog No. No. of Reactions Includes
A39005096 96 Reactions RNA Prep, Depletion, UD Index
A38994024 24 Reactions RNA Prep Kit Only
A38994096 96 Reactions RNA Prep Kit Only
A39003024 24 Reactions RNA Prep, Depletion Kit
A39003096 96 Reactions RNA Prep, Depletion Kit
A39005024 24 Reactions RNA Prep, Depletion, UD Index
A38996024 24 Reactions RNA Prep and UD Index
A38996096 96 Reactions RNA Prep and UD Index
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Catalog No. A39005096 Supplier Invitrogen™ Supplier No. A39005096

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

Kits are designed for next-generation sequencing (NGS) on Illumina systems and support whole-transcriptome sequencing (human, mouse, rat), and mRNA sequencing (compatible with all species) applications.

The Invitrogen Collibri Stranded RNA Library Preparation kits are designed for next-generation sequencing (NGS) on Illumina systems. The kit supports whole-transcriptome sequencing (human, mouse, rat) and mRNA sequencing (compatible with all species) applications.

The Invitrogen Collibri Stranded RNA Library Preparation kits are designed for next-generation sequencing (NGS) on Illumina systems. The kit supports whole-transcriptome sequencing (human, mouse, rat) and mRNA sequencing (compatible with all species) applications.

The Collibri Stranded RNA Library Preparation Kit:
• Includes all necessary components in a single kit* for a quick use
• Total RNA converted to sequencing-ready library within 4.5 hours
• >98% strand specificity, efficient detection of non-coding RNA and high sensitivity of differential gene expression
• Provides in-process visual feedback for easier handling
• Is suitable for conventional automation to increase throughput

The Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina Systems is designed for robust construction of cDNA libraries for strand-specific RNA sequencing on Illumina next-generation sequencing (NGS) systems. The kit brings together the superior features of SuperScript IV Reverse Transcriptase, Dynabeads magnetic particles, and Platinum SuperFi DNA Polymerase, enabling users to obtain high-quality sequencing-ready libraries. This Collibri Stranded RNA Library Prep Kit supports fast library preparation within 4.5 hours for mRNA or whole transcriptome libraries with 1–25 ng of mRNA-enriched or ribodepleted RNA sample input. Multiplexing of libraries can be carried out using up to 96 single-indexed or unique dual-indexed primers, enabling single-read or paired-end sequencing. The kit is suitable for various-quality RNA samples, including FFPE samples.

Visual cues of process integrity
For maximum convenience, visual cues of process integrity are included throughout the library preparation workflow. Monitor progress of library generation using visual feedback from dyes in critical reagents. The reaction mix changes color in response to the addition of critical components at each step to ensure that library preparation has the highest chance of success. The inert dyes do not interfere with enzymatic reactions and do not compromise sequencing results.

How it works
Full-length Illumina-compatible adaptors are introduced by PCR amplification using Platinum SuperFi DNA Polymerase to construct single-indexed or unique dual-indexed libraries compatible with single-read or paired-end sequencing. The kit contains 24 or 96 either single-indexed barcoded PCR primers (i7) pre-mixed with a universal i5 primer or unique dual-indexed PCR primers that enable multiplexing of up to 96 libraries. Optimized cleanup steps efficiently remove residual primers and adapter/primer dimers while preserving high library yields.

A related product, the Collibri Library Quantification Kit, is recommended for qPCR-based quantifications of libraries before proceeding to sequencing.

Applications
The Collibri Stranded RNA Library Prep Kit is recommended for:
• Gene expression studies
• Alternative splicing analysis
• Non-coding RNA detection and discovery
• Identification of alternative polyadenylation sites
• Gene fusion detection
• Detection of transcript isoforms

* The kit consists of enzymatic modification, size selection/clean-up, and indexed adapter-containing modules. Additional ribodepletion module of human/mouse/rat rRNA can also be included.

Specifications

Content And Storage • Collibri Library Prep Kit, store at -20°C
• Collibri Library Cleanup Kit, store at 4°C
Description The product contains modules A39117096, A39119196, A39115096, A39114096
Sample Type RNA
For Use With (Equipment) Illumina NGS Instruments
Includes RNA Prep, Depletion, UD Index
No. of Reactions 96 Reactions
Libraries cDNA Library
Product Line Collibri
Quantity 96 reactions
Sequencing Type Transcriptome Sequencing
Shipping Condition Approved for shipment on Wet or Dry Ice
Workflow Step Library Generation
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Frequently Asked Questions (FAQs)

Is the elution buffer, included in the Collibri Stranded RNA Library Prep Kit for Illumina Systems, "Tris buffered" or "Tris-Tween"?

Collibri Stranded RNA Library Prep Kit for Illumina Systems elution buffer is based on Tris-HCl, with no EDTA or EGTA.

Is there an RNA control for optimizing fragment length included with the Collibri Stranded RNA Library Prep Kit for Illumina Systems?

The Collibri Stranded RNA Library Prep Kit for Illumina Systems does not include an RNA control. Recommendations for RNA controls can be found in the Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina User Guide, page 16 "Guidelines for RNA Controls".

What are the recommended RNA input sample types and respective sample amounts for Collibri Stranded RNA Library Prep Kit for Illumina Systems?

Information for RNA sample type and amount can be found in the Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina User Guide, page 15 "Guidelines for RNA sample type and amount".

What are the differences between all Collibri Stranded RNA Library Prep Kit for Illumina Systems catalog numbers?

Information regarding the different catalog numbers can be found in the Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina User Guide, page 3.
The different catalog numbers allow for the option to purchase different sizes of the library kit and with or without Unique Dual Indexes.

What additional materials are required when using Collibri Stranded RNA Library Prep Kit for Illumina Systems?

A list of required materials that are not supplied with the kit can be found in the Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina User Guide, page 11-12.

What is the difference between the Collibri Stranded RNA Library Prep Kit for Illumina Systems Cat. Nos. A38994024 and A38996024?

Unlike Cat. No. A38994024, Cat. No. A38996024 comes with the Unique Dual Indexes (UDIs), listed in table 6, page 10 of the Invitrogen Collibri Stranded RNA Library Prep Kit for Illumina User Guide

Please also refer to the Technical Note "How to prevent index hopping" for additional information.

In general, UDIs have a sample index that is specific to a given library. All nucleic acids in a sample are labeled with the same sequence tag and the resulting library is pooled with other libraries and sequenced in parallel in a single run. UDIs are the preferred method when multiplexing to mitigate sample misassignment due to index hopping and the reagent kits supply enough to perform extra cycles. UDIs require extra cycles and the reagent kits supply enough to perform those extra cycles. UDIs can be especially effective when using instruments with patterned flow cells, such as the NovaSeq 6000 system.

Can indexed adaptors be used interchangeably between RNA-seq (RNA sequencing) and DNA-seq (DNA sequencing)?

No, indexed adaptors are specifically designed for each product.
Are adaptors included in the Collibri RNA library prep kits?

Yes. Ready-to-ligate adaptors are included in the Collibri RNA library prep kits. Please note that the adaptors are not available separately and are provided only as an integral part of the Collibri kits.
With the Collibri RNA library prep kits, when should the ERCC RNA Spike-In Control Mixes be added?

ERCC RNA Spike-In Control Mixes should be added to the sample before all other steps and should undergo exactly the same treatment as sample RNA. For whole transcriptome library generation, we recommend that you add ERCC RNA Spike-In Control Mixes to the input RNA before rRNA depletion or poly (A) selection.
With the Collibri RNA library prep kits, what is the advantage(s) of performing the cDNA step after the adaptor ligation?

Once the 5' and 3' adaptors are ligated to the RNA, cDNA synthesis starts not from random hexamers, but from one universal oligo that binds to the 3' adaptor. This eliminates bias from the priming site and also ensures conversion of every fragment into cDNA: random hexamers have to find a landing spot and some fragments do not have it and do not make it into the library. It also eliminates random nucleotide-introduced mistakes: during second-strand synthesis, a random hexamer becomes a legitimate part of the library, introducing “SNPs”, “point mutations” etc. These events are filtered out by software but nevertheless, decrease the output quality. With Collibri RNA library prep kit, adaptors never become part of the library.
Can the Collibri RNA library prep kits be adapted for automation?

Yes, the Collibri RNA library prep kits can be adapted for automation.

For Research Use Only. Not for use in diagnostic procedures.