Collibri™ Stranded RNA Library Prep Kit for Illumina™ Systems with Human/Mouse/Rat rRNA Depletion Kit, 24 rxns - FAQs

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.

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.

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.

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.

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.

No, indexed adaptors are specifically designed for each product.

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.

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.

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.

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