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  • Cappable-seq Service now available !

    In close cooperation with New England Biolabs, Inc. (NEB), VERTIS is now able to make the new Cappable-seq method available to its customers. The method was developed by Laurence Ettwiller and Ira Schildkraut from NEB (Ettwiller, L. et al. 2016. BMC Genomics. 17, 1 (2016), 199).

    It represents the current most sensitive and robust method for the precise identification of bacterial transcription start sites (TSSs).

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WTSS FL-cDNA Normalization MicroRNA PARE DGE

cDNA Synthesis in full length

We offer the preparation of full-length cDNA libraries. The libraries are best suited for isoform sequencing (Iso-Seq) using the PacBio RS II system, which enables sequencing of transcripts from the 5‘ end to the poly(A) tail.

In order to obtain optimum sequence coverage from a PacBio sequence run, gene representation in the libraries can be equalized using vertis normalization technology.

We use different strategies for the preparation of FL-cDNA using oligo-dT priming of total or poly(A)+ RNA or a combination of oligo-dT and random-priming using rRNA depleted RNA for 1.-strand cDNA synthesis. The different methods are outlined below.

 

Method 1

Method 2

Method 3

Adatper ligation to 1.-strand cDNA

Starting material:
Total or poly(A)+ RNA

Priming: oligo-dT

Advantage of method:
Robust, good coverage also of transcripts which are not 100% full-length 

Oligo ligation to 5' ends of RNA

Starting material:
Total or poly(A)+ RNA

Priming: oligo-dT

Advantage of method:
Possibility to enrich for true full-length transcripts

Oligo ligation to 5' ends of RNA

Starting material:
ribo-depleted RNA

Priming: oligo-dT + random N6

Advantage of method:
Possibility to also capture transcripts without poly(A) tails

 

 

 

 
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