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Collibri™ Stranded RNA Library Prep Kit for Illumina™ Systems, 24 preps - FAQs

View additional product information for Collibri™ Stranded RNA Library Prep Kit for Illumina™ Systems - FAQs (A39005024, A39003024, A38996096, A39003096, A38994096, A39005096, A38996024, A38994024)

11 product FAQs found

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.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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".

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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".

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

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.