Illumina

Bacterial community amplicon sequencing

• Multiple Amplicon targets

• 16S rRNA gene sequencing V4 (multiple versions).

• We have validated studies with other primer targets. Please ask if your target-of-interest could be used for bacterial community projects.

Fungal community amplicon sequencing

• Multiple Amplicon targets

• ITS1-2

• We have validated studies with other primer targets, so please ask if your target-of-interest could be used for fungal community projects.

Small genome sequencing

• Bacteria

• Nextera XT

• Eukaryotic viruses

• Nextera XT

• Bacteriophages

• Nextera XT

Shotgun Metagenomics

• Untargeted

• Viral metagenomics DNA and RNA viruses.

• Methods and bioinformatics designed by academic team.

Bacterial transcriptomics

• Using ScritpSeq Library Preparation sequencing kit

• https://emea.illumina.com/products/scriptseq-rna-seq-library-prep.html

TruSeq, panel/design for eukaryotic gene expression

Contact for further details or follow the below link to find validated Illumina panels.

https://emea.illumina.com/products/selection-tools/gene-panel-finder.html#/targeted-panels

miRNA sequencing

• TruSeq Small RNA sequencing

• http://emea.support.illumina.com/content/dam/illumina-support/courses/TruSeq_Small_RNA/index.html?iframe

• http://emea.support.illumina.com/content/dam/illumina-support/documents/documentation/chemistry_documentation/samplepreps_truseq/truseqsmallrna/truseq-small-rna-library-prep-kit-reference-guide-15004197-02.pdf

Please send samples to the following delivery address;

NU-OMICS DNA sequencing research facility
(C/O) Darren Smith/Andrew Nelson
Northumbria University
Ellison Building EBA 504a
Newcastle Upon Tyne
Tyne and Wear
NE1 8ST

Small Genome sequencing; Bacterial or virus

• NexteraXT

• All samples must be treated with broad spectrum RNAse prior to sending. An electrophoresis image is required for QA purposes run on a 1% agarose gel. Limited shearing should be present, if worried contact us to check before sending.

• All samples should be in a 96 well format as parts of the preparation are automated. Data will be returned labelled in well-format over sample name. DNA quality A260/280 should be >1.8.

• For Nextera XT we require a minimum of 55 ng of DNA in 11ul of elution buffer.

Bacterial and Fungal Community amplicon sequencing

• DNA sample extraction should be with either phenol:chloroform or for kit based methods we recommend the Qiagen (MoBio) PowerSoil DNA extraction kit.

• All samples should be in a 96 well format as parts of the preparation are automated. Data will be returned labelled in well-format over sample name. DNA quality A260/280 should be >1.8. Please trial the DNA extraction to see if possible to amplify the target gene in a subset of your samples.

• Always include a DNA extraction negative. If you use an extraction kit or require multiple kits due to sample numbers please provide a DNA extraction negative for each kit. We will provide a sequencing negative and a commercial mock community so that downstream analysis can be completed (https://www.zymoresearch.eu/zymobiomics-community-standard)

Bacterial RNA sequencing

• Using ScritpSeq Library Preparation sequencing kit

• We require a minimum of 1ug of total RNA with RIN value >1.8, although higher is recommended. We can provide QA of total RNA extraction at an extra cost.

• We are happy to accept libraries already prepared for pooling as long as the supporting QA is available.

Shotgun metagenomics

• NexteraXT

• All samples must be treated with broad spectrum RNAse prior to sending. An electrophoresis image is required for QA purposes run on a 1% agarose gel. Limited shearing should be present, if worried email Nuomics@northumbria.ac.uk to check before sending.

• All samples should be in a 96 well format as parts of the preparation are automated. Data will be returned labelled in well-format over sample name. DNA quality A260/280 should be >1.8.

• For Nextera XT we require a minimum of 55 ng of DNA in 11ul of elution buffer.

Targeted RNA sequencing from predesigned or bespoke amplicon panels (TruSeq)

• We require a minimum of 1ug of total RNA with RIN value >1.8, although higher is recommended.

• We are happy to accept libraries already prepared for pooling as long as the supporting QA is available.

Small RNA sequencing

• We require a minimum of 1ug of total RNA with RIN value >1.8, although higher is recommended.