Project Budget (Detailed)
Detailed Project Budget
All monetary values are by default in CAD unless stated otherwise.
Stage 1 - Cell Culturing
Killifish Growth Medium
Base medium taken from: https://doi.org/10.1016/j.cbpa.2014.05.002
Added IGF, insulin, bFGF
Component | Cost |
90.50 (500 mL) | |
960.00 (500 mL) | |
22.98 (20mL) | |
+/- insulin-like growth factor 1 (IGF-1, Shenandoah Biotechnology #100-34) | 101.02 USD => 137.00 CAD (for 20 μg) |
Insulin solution from bovine pancreas (Sigma Aldrich #I0516) | 137.00 (for 5 mL) |
623.29 (for 10μg) | |
Total | 1970 |
Pricing information as of October 4, 2023.
Stage 2 - Library Preparation
We are not requesting funding for this stage of the project. We’re confident that the combination of the funds we currently have available along with funding from other sources will be able to cover this stage.
Price quotes are from McMaster University’s Genomics facility. They’re one of the few facilities in Canada that offer both scRNA and scATAC sequencing. Library preparation of the samples must be done prior to actual sequencing.
Library Preparation
Item | Amount | Per item cost | Total |
10x SC Multiome ATAC + GX library prep, per batch (≤8 samples) | 1 | 1,550.00 | 1,550.00 |
10x SC Multiome ATAC + GX library prep, per sample | 6 | 5,350.00 | 32,100.00 |
Illumina library QC (per batch, ≤11) | 1 | 265.00 | 265.00 |
Illumina library QC (per sample) | 6 | 14.00 | 84.00 |
Illumina library QC, small batch (per batch, ≤6) | 1 | 150.00 | 150.00 |
Illumina library QC, small batch (per sample) | 1 | 25.00 | 25.00 |
Total | 34,174 |
Note: GX stands for Gene Expression (RNA). 10x SC Multiome ATAC + GX library allows for joint scRNA/scATAC-seq.
Stage 3 - scRNA/scATAC sequencing
Reagant Kits
A P1 kit has 100M reads. Cost: 1,850.00
A P2 kit has 400M reads. Cost: 2,800.00
A P3 kit has 1100M reads. Cost: 6,000.00
Gene expression (RNA) libraries require a minimum of 20k reads per cell and ATAC libraries require a minimum of 25k reads per cell. The number of cells that we can input per sample is 500-10000. So for example, if we wanted to sequence 3000 cells per sample x 18 samples, we would need 1080M reads at a minimum, which would require a P3 kit which has 1100M reads. Each type of library must be sequenced separately. Even if you go ahead with the Multiome ATAC + Gene expression kit, that would still be one NextSeq kit for the ATAC sequencing, and another kit for the gene expression sequencing (normal single cell).
Both types of libraries would be run on the NextSeq 2000 using a 2x50 configuration.
Number of cells per sample: 6000
Number of reads per cell: 30000
Number of samples: 6
We have 3 samples per experiment (one at each of the 3 time points)
We are running 2 experiments to ensure reliability: 2 x 3 = 6
scRNA: 6 samples x 6000 cells/sample x 30000 reads/cell = 1080M reads (1 x P3 kit)
scATAC: 6 samples x 6000 cells/sample x 30000 reads/cell = 1080M reads (1 x P3 kit)
2 x P3 kit = 2 x $6000 = 12000
Total: 12000.00
Final Total: $48,144 CAD => $35,070 USD
Amount Requested: 1970 + 12000 = $13970 CAD => $10,175 USD
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