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Research and Innovation

RNA sequencing

Gene expression profiling for Eukaryotes

RNA samples are prepared using a strand-specific mRNA sample preparation kit using poly(A) isolation with multiplexing.  ERCC RNA controls are added prior to library preparation providing additional control for variability.  RNA library quality, quantity and insert size is assessed by a combination of Tapestation analysis, Qubit fluorescence assay and qPCR prior to sequencing.  
 
What we offer: 
Sample receipt and addition of ERCC RNA control  
Poly(A) selection      
Stranded cDNA Synthesis and multiplexed library generation 
Illumina Sequencing (2x 151bp Reads) 
20 million, 30 million or 50 million reads per sample 
Standard Illumina QC   and  Bioinformatics Analysis for RNA-seq 

input requirements:

  • Sample Type: Purified total RNA (DNA-free)
  • Recommended RIN: ≥8.0 (For RIN value ≥ 5.5 contact us)
  • Recommended Quantity: Samples normalised between 1 ng/μl to 40 ng/μl. The higher the concentration, the lower the number of PCR cycles performed. 
  • Volume: 55 μl
  • Resuspension Buffer: Nuclease-free water
  • Shipment Method: Dry ice, overnight  
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume  

Total RNA sequencing for Prokaryotes, Eukaryotes or Metatranscriptomics

RNA samples are prepared using rRNA depletion, suitable for prokaryotes and eukaryotes, including depletion of globin transcripts if samples have been isolated from blood. ERCC RNA controls are added prior to library preparation providing additional control for variability. Libraries are strand-specific. UMI’s can be added if required.  RNA library quality, quantity and insert size is assessed by a combination of Tapestation analysis, Qubit fluorescence assay and qPCR prior to sequencing. 
 
What we offer 
Sample receipt and addition of ERCC RNA control  
Globin/rRNA depletion      
Stranded cDNA Synthesis and multiplexed library generation 
Illumina Sequencing (2x 151bp Reads) 
10 million 30 million, 50 million or 100 million reads per sample 
Standard Illumina QC   and Transcriptome generation 

Input requirements rRNA removal (for Prokaryotes and Eukaryotes)

  • Sample Type: Purified total RNA (DNA-free) 
  • Recommended RIN: ≥8.0
  • Recommended Quantity: Samples normalised between 1 ng/μl to 91 ng/μl. The higher the concentration, the lower the number of PCR cycles performed.
  • Volume: 30 μl 
  • Resuspension Buffer: Nuclease-free water
  • Shipment Method: Dry ice, overnight 
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume, Sample RIN 

Full length transcriptome sequencing utilising Nanopore technology

This method is beneficial for identifying and quantifying full length transcripts and can reduce multiple locus alignment issues. It is provides clarity when characterising isoforms, splice variants and fusion transcripts. ERCC RNA controls can be added prior to library preparation if required. The polyA-RNA is reverse transcribed, strand-switched and a UMI is incorporated before PCR amplification and barcoding. Finally, rapid sequencing adapters are added.  For human samples we would aim for 20 million reads /sample. 
 
What we offer 
Sample receipt and QC  
cDNA Synthesis from oligoDT primer and multiplexed library generation using cDNA PCR barcoding kit using kit 14 chemistry (24 barcodes available) 
Sequencing full length transcripts on a Promethion generating 50-180Gb per flowcell (suitable for 3-6 human transcriptomes) 
20 million reads per sample for humans, other options are available. 
Standard QC Nanopore   Do we have some Nanopore transcriptome analysis that we offer? 

Input requirements 

  • Sample Type: Total RNA or polyA
  • Sample Purity (OD260/OD280): 1.8-2.2
  • Recommended RIN: ≥7.0 
  • Total RNA 
    • Recommended Quantity: ≥500 ng, >50 ng/µl
    • Minimum Quantity: 250 ng, 20 ng/ul 
  • PolyA 
    • Recommended Quantity: ≥10 ng, >1 ng/µl
    • Minimum Quantity: 5 ng, 0.5 ng/ul
  • Resuspension Buffer: Nuclease-free water
  • Shipment Method: Dry ice, overnight 
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume, Sample RIN 

Small RNA

Small non-coding RNAs are involved in gene silencing and post-transcriptional modification of coding RNAs. The molecules are between 20 and 200 based in length and include microRNA, piRNA, snRNA, viral RNA etc. Profiling small RNAs increases understanding of how post-transcriptional RNA modification contributes to phenotype and enables identification of bio markers. When planning a small RNA experiment it is important to ensure the method of RNA isolation does not exclude these small molecules. 
 
What we offer 
Sample receipt and QC 
Small RNA library generation and size selection 
Illumina Sequencing (2x 151bp Reads) 
10 million reads per sample 
 
Input recommendations 

  • Sample Type: Total RNA (containing small RNA and DNA-free)
  • Recommended Quantity: Samples normalised between 1 ng/μl to 400 ng/μl, ideally 200 ng/μl. The higher the concentration, the lower the number of PCR cycles performed. 
  • Recommended RIN: ≥7.0  
  • Volume: 20 μl
  • Resuspension Buffer: Nuclease-free water
  • Shipment Method: Dry ice, overnight
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume, Sample RIN 

Circular RNA

Circular RNAs are highly stable non-coding RNA that have a role in gene regulation ..... 
 
Input recommendations 

  • Sample Type: Total RNA (containing DNA-free circular RNA)
  • Recommended Quantity: Samples normalised between 100 ng/μl to 400 ng/μl, ideally 200 ng/μl. The higher the concentration, the lower the number of PCR cycles performed.
  • Recommended RIN: ≥8.0  
  • Volume: 50 μl
  • Resuspension Buffer: Nuclease-free water
  • Shipment Method: Dry ice, overnight 
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume, Sample RIN 

RNA samples are prepared using a strand-specific mRNA sample preparation kit using poly(A) isolation with multiplexing.  ERCC RNA controls are added prior to library preparation providing additional control for variability.  RNA library quality, quantity and insert size is assessed by a combination of Tapestation analysis, Qubit fluorescence assay and qPCR prior to sequencing.  

What we offer: 

Sample receipt and addition of ERCC RNA control  

Poly(A) selection      

Stranded cDNA Synthesis and multiplexed library generation 

Illumina Sequencing (2x 151bp Reads) 

20 million, 30 million or 50 million reads per sample 

Standard Illumina QC   and  Bioinformatics Analysis for RNA-seq 

input requirements: 

  • Sample Type: Purified total RNA (DNA-free) 
  • Recommended RIN: ≥8.0 (For RIN value ≥ 5.5 contact us) 
  • Recommended Quantity: Samples normalised between 1 ng/μl to 40 ng/μl. The higher the concentration, the lower the number of PCR cycles performed. 
  • Volume: 55 μl 
  • Resuspension Buffer: Nuclease-free water 
  • Shipment Method: Dry ice, overnight  
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume  

RNA samples are prepared using rRNA depletion, suitable for prokaryotes and eukaryotes, including depletion of globin transcripts if samples have been isolated from blood. ERCC RNA controls are added prior to library preparation providing additional control for variability. Libraries are strand-specific. UMI’s can be added if required.  RNA library quality, quantity and insert size is assessed by a combination of Tapestation analysis, Qubit fluorescence assay and qPCR prior to sequencing. 

What we offer 

Sample receipt and addition of ERCC RNA control  

Globin/rRNA depletion      

Stranded cDNA Synthesis and multiplexed library generation 

Illumina Sequencing (2x 151bp Reads) 

10 million 30 million, 50 million or 100 million reads per sample 

Standard Illumina QC   and Transcriptome generation 

Input requirements rRNA removal (for Prokaryotes and Eukaryotes)  

  • Sample Type: Purified total RNA (DNA-free) 
  • Recommended RIN: ≥8.0 
  • Recommended Quantity: Samples normalised between 1 ng/μl to 91 ng/μl. The higher the concentration, the lower the number of PCR cycles performed. 
  • Volume: 30 μl 
  • Resuspension Buffer: Nuclease-free water 
  • Shipment Method: Dry ice, overnight 
  • Minimum Sample data: including; project ID, sample ID, sample concentration, sample volume, Sample RIN