We are offering end to end services from extraction to sequencing or modular individual services, we can customize projects to fit individual needs.
- Extractions (DNA or RNA)
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Health Sciences Sequencing Core
Sample Type
Input Requirements*
Price
Cells
0.5-10 million cells
$32.00
Tissue
5-30mg tissue
$32.00
Paraffin-embedded (FFPE) Tissue
Please provide three 5mm sections
$47.00
Saliva
Sample provided in a DNAGENOTEK Saliva-based collection tube (www.dnagenotek.com)
$47.00
Blood/Serum/Plasma/Buffy Coat
Sample provided in:
1. BD Vacutainer Blood Collection tube (www.bd.com)
2. Paxgene tube
3. Tempus tube
Note: please follow manufacturer protocol/recommendations for each tube type
$47.00
*Input requirements are recommended. If your sample amount is limited, contact us to discuss options.
- Input Requirements
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RNA Library preparation
Input Quantity range
RNA Integrity
(RIN) Score
Min Concentration
Illumina Stranded Total Ligation (Ribo-Zero Plus)
10-1000 ng
RIN > 2
1 ng/µl
Illumina Stranded mRNA (PolyA+)
25-1000 ng
RIN ≥ 8
1 ng/µl
SMARTer TCR Profiling (Human / mouse)
10-250 ng
RIN > 7
1 ng/µl
SMART-Seq Stranded Total RNA (Ultra Low Input)
1-10 ng
RIN > 2
0.13 ng/µl
SMART-Seq HT Ultra Low Input
1-10 ng
RIN ≥ 8
0.2 ng/µl
QIAseq small RNA
1-500 ng
RIN ≥ 8
0.2 ng/µl
DNA Library preparation
Nextera DNA XT (low input)
1 ng cDNA
0.2 ng/µl
Illumina DNA Prep (Nextera FLEX)
1-500 ng cDNA
0.2 ng/µl
Illumina DNA PCR-Free Prep (WGS)
25-300 ng DNA
1 ng/µl
ChIP-seq (Library prep only)
5-10 ng CHIP DNA
0.1 ng/µl
ATAC-Seq
10k-50k cells
- Library Generation
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lllumina Stranded Total Ligation (Ribo-Zero Plus)
Total RNA input is depleted using enzymatic depletion method that removes abundant transcripts from total RNA. Random primers initiate first and second strand cDNA synthesis. Adenylation of 3’ ends are followed by adapter ligation and library amplification with indexing.
This method is used when you want to characterize long non-coding RNA (lncRNA) as well as mRNA. It is compatible with degraded RNA as there is no poly-A enrichment.
lllumina Stranded mRNA (PolyA+)
Total RNA input is enriched for mRNA molecules containing polyA tails using Oligo(dT) magnetic beads. The purified mRNA is fragmented, and Random primers initiate first strand and second strand cDNA synthesis. Adenylation of 3’ ends are followed by adapter ligation and library amplification with indexing.
This method is used in the transcriptome or gene expression studies.
SMARTer TCR Profiling (Human / mouse)
It is used to analyze T-cell receptor (TCR) diversity from total (bulk) RNA samples and whole cells. cDNA is synthesis using TCR dT primes. Two rounds of PCR are performed to amplify cDNA sequences. In second PCR, forward and reverse primers is added including adapter and index sequence.
SMART-Seq Stranded for Total RNAseq (Ultra Low Input)
Reverse transcription with random primers and template switching generates first strand cDNA. cDNA is amplified using the template switching adapter and a sequence added to the random primers. Ribosomal cDNA is cleaved and the uncleaved fragments are amplified with indexed primers. This method is used with low quality and quantity RNA samples.
SMART-Seq mRNA HT (Ultra Low Input) – for flow sorted cells
SMART-Seq HT ultra-Low input RNA Kit – prepare ds-cDNA
Reverse transcription with oligo(dT) primers and template switching oligo generates first strand cDNA. cDNA is amplified using the template switching adapter and a sequence added to the polyT primer.
This kit is used for very low input RNA and/or direct flow sorted samples.
Illumina Nextera XT
Tagmentation with transposase fragments DNA and incorporates shortened Illumina adapters. Indexes are added via index PCR.
Illumina DNA Prep (previously named: Nextera FLEX)
Tagmentation using enzymatic reaction to fragment DNA and add adapter sequences. Indexes are added via index PCR.
Illumina DNA PCR-free
Tagmentation using enzymatic reaction to fragment DNA and add adapter sequences. Indexes are added via direct ligation. Used for whole genome sequencing (WGS).
Lexogen 3’-Quantseq
QuantSeq kits enable cost-efficient sequencing by counting (differential gene expression only). These kits are an alternative to standard Total RNA-Seq from highly degraded low-input samples. Library is generated from 3’-ends only. Not recommended for lncRNA and isotype variants.
QIAseq small RNA
Pre-adenylated adapter is ligated to the 3’ end of small RNAs followed by 5’ ligation of an RNA adapter. cDNA synthesis is completed using a reverse transcription (RT) primer targeted to the 3’ pre-adenylated adapter. Unique molecular indices (UMIs) are integrated into the RT primer to assign a unique identifier to each miRNA molecule. Indexes are added via index PCR.
ChIP-seq (Library prep only)
The goal of this protocol is to add adapter sequences onto the ends of ChIP DNA to generate indexed libraries.
ATAC-Seq/Omni-ATAC-seq
Assay for transposase-accessible chromatin sequencing to identify regions of open chromatin. in the tagmentation process, Tn5 transposase fragments open genomic DNA and inserts sequencing primers into chromatin open regions. Data analysis from sequencing provide understanding of gene regulation.
- Illumina Sequencing
-
Health Sciences Sequencing Core
Miseq Flow Cells
Number of Reads
Cycles
Typical Run type
Price
Micro
4 million
300
2x150bp
$620
V2
13 million
50
1x50bp
$1180
300
2x150bp
$1460
V3
25 million
150
2x75bp
$1280
600
2x300bp
$2050
NextSeq 2000 Flow Cells
Number of Reads
Cycles
Typical Run type
Price
P1
100 million
100
2x58bp
$1300
600
2x300bp
P2
400 million
100
2x58bp
$1900
200
2x100p
$3450
300
2x150bp
$4480
P3
1.2 billion
50
1x50bp, 2x35bp
$3100
100
2x58bp
$4000
200
2x100bp
$5600
300
2x150bp
$7350
- Single Cell Technologies
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3’ Gene Expression
Gene expression of up to 10,000 cells per sample. Multiplexing options include cell hashing with antibody labelling or CellPlex lipid tags.5’ Gene Expression and immune profiling
Gene expression, TCR and BCR profiling of up to 10,000 cells per sample. Multiplexing available with antibody-based cell hashing.Gene Expression Flex
This is a probe-based assay that can be used to profile fixed tissues and cells (included cells isolated from FFPE). Tissues and cells can be stored up to 6 months at -80C. Multiplexing available for up to 16 samples. Compatible with human and mouse.CITE-Seq
Barcoded antibodies bind to surface markers on individual cells, the barcode marker sequence carries over through the library preparation, permitting quantitative assessment of surface antibody binding to cells along with their transcriptome information. Hundreds of surface antigens can be assayed simultaneously.Multiome ATAC and Gene Expression
Provides simultaneous detection of mRNA and chromatin accessibility from the same cell.For more information, please visit https://singlecell.pitt.edu/
- Spatial Transcriptomics
-
Health Sciences Sequencing Core
10X Visium Spatial Gene Expression
For additional information: https://www.10xgenomics.com/products/spatial-gene-expression
Sample Type
Tissue size
Samples per slide
Staining Options
Cost
(Sequencing not included)
Fresh Frozen
6.5mm x 6.5mm
4
H&E, Immunofluorescence (4 colors max)
$6,200
+ $1,200 for tissue permeabilization (required)
CytAssist Fresh Frozen
6.5mm x 6.5mm
4
H&E only
$8,400
CytAssist Fresh Frozen
11mm x 11mm
2
H&E only
$8,600
CytAssist
FFPE
6.5mm x 6.5mm
4
H&E, Immunofluorescence (4 colors max)
$8,400
CytAssist
FFPE
11mm x 11mm
2
H&E, Immunofluorescence (4 colors max)
$8,600
Curio Biosciences Slide Seeker
For additional information: https://curiobioscience.com/
Sample Type
Tissue size
Samples per slide
Staining Options
Cost
Fresh Frozen
3mm x 3mm
1
None for transcriptome section – serial sections for staining recommended
Please contact
Fresh Frozen
10mm x 10mm
1
None for transcriptome section – serial sections for staining recommended
Please contact