NovaSeq 6000 Cartridge - Sequencing

RNA and epigenetic sequencing

The iGenSeq platform at Paris proposes the RNA sequencing of human, murine, plant or bacterial origin. Thanks to the experience acquired since 2011 in this technique, a wide range of library preparations is available to answer a wide variety of biological questions:

Messenger RNA sequencing (opens a new tab)

Several kits are available for sequencing mRNAs according to the quantity of RNA available. The KAPA mRNA HyperPrep kit (Roche) is used routinely. For smaller quantities, and for your specific requests, we offer a wider range of kits (see table below).

 

Targeted RNAs kit Quantity Volume Quality controls to be done by the customer
mRNA KAPA mRNA HyperPrep (Roche) 50 ng to 1µg of total RNA 50 µl Qubit / Nanodrop + Bioanalyzer
mRNA Stranded mRNA (Illumina) 25 ng to 1μg of total RNA 25 µl Qubit / Nanodrop + Bioanalyzer
mRNA Ultra II Directional RNA library (NEB) 10 to 1 µg of total RNA 50 µl Qubit / Nanodrop + Bioanalyzer
mRNA Smartseq V4 (Takara) + Nextera XT (Illumina) 10 pg to 10 ng total RNA 9,5 µl max Qubit / Nanodrop + Bioanalyzer

 

Sequencing of degraded RNA or from FFPE tissues

For the study of RNAs whose quality is compromised, in particular those resulting from laser microdissection or FFPE tissues, iGenSeq offers two types of service, depending on the quality of the material available:

Targeted RNAs kit Quantity Volume Quality controls to be done by the customer
Degraded RNA QuantSeq 3 ′ mRNA-Seq Lib Prep Kit FWD (Lexogen) 300 ng 10µL Qubit / Nanodrop + Bioanalyzer
Degraded RNA SMARTer Stranded Total RNA-Seq Kit v3 - Pico Input Mammalian (Takara) 0,25 to 10 ng 8µL Qubit / Nanodrop + Bioanalyzer

 

RNA sequencing with globin and ribosome depletion

Samples from blood samples are abundantly rich in globin RNA. To study the RNAs obtained from these samples, it is essential to remove this type of messenger RNA before preparing the libraries for these samples. The platform offers this service coupled with the depletion of ribosomal RNAs for the study of long non-coding RNAs that do not have a polyadenylated tail:

 

Targeted RNAs kit Quantity Volume Quality controls to be done by the customer
Total RNA except globin and ribosome RNA Ribo-Zero Globin + TruSeq Stranded Total RNA (Illumina) 1 ng to 1 μg total RNA 11 µl max Qubit / Nanodrop + Bioanalyzer

 

Small RNA sequencing

To study the small RNAs involved in the post-transcriptional regulation of gene expression, we offer the following kits depending on the origin of the sample:

Targeted RNAs kit Quantity Volume Quality controls to be done by the customer
microRNA (cells; tissues) Qiaseq miRNA Library (Qiagen) 1 ng to 500 ng total RNA 5 µl Qubit / Nanodrop + Bioanalyzer
microRNA (serum; plasma) Qiaseq miRNA Library (Qiagen) * 5 µl of RNA ** 5 µl Qubit / Nanodrop + Bioanalyzer

*Use the extraction kit compatible with miRNeasy Serum / Plasma plasma / serum samples

** Elute 5 µL of RNA from 200 µl of serum or plasma

 

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