Instrumentation and Services
Core Provide training for all shared instruments, and they are operated under Self Service. Please use this link https://ppms.us/salk/listsys/?pf=9 to book those instruments on PPMS
Agilent TapeStation 4150 for measuring DNA
| Nucleic Acid | Tape Station Assay | Description | Concentration Range (ng/μl) | Size Range (bp) |
|---|---|---|---|---|
| RNA | RNA Tape | RNA Screen Tape | 5-500 | 50-600 |
| HS RNA Tape | High Sensitivity RNA Screen Tape | 0.1-10 | 50-600 | |
| DNA | DNA Tape | D1000/D5000 Screen Tape | 0.1-50 | 35-1000 |
| HS DNA Tape | High Sensitivity D1000/D5000 Screen Tape | 0.01-1 | 35-1000 | |
| Genomic Tape | Genomic DNA Screen Tape | 10-100 | 200-15000 |
Qubit 3.0 Fluorometer for measure DNA or RNA concentration with all the reagents provided. Remember to sign out the reagents usages when finished.
| Nucleic Acid | Qubit Assay | Description | Concentration Range (ng/μl) | Sample Types |
|---|---|---|---|---|
| RNA | BR RNA | Qubit Broad Range RNA | 5-1000 | RNA |
| HS RNA | Qubit High Sensitivity RNA | 0.25-100 | RNA from limited samples | |
| DNA | BR DNA | Qubit Broad Range DNA | 0.1-1000 | Genomic, dsDNA |
| HS DNA | Qubit High Sensitivity DNA | 0.1-100 | dsDNA, ChIP |
QuantStudio 3 and 5 Real-Time PCR Systems can detect differences in target quantity as low as 1.5-fold, support 96-well (0.2ml or 0.1ml) or 384-well (QS5 only) consumables. The recharge is linked to the time it was booked.
Tecan Infinite® 200 Plate reader for measuring fluorescence, luminescence or absorbance detection with an advanced quad-4 monochromators™ system (M200). It has the i-control™ software for easy set up protocols, and optional Magellan™ software packages provide full data reduction capability. It is no charge for using it if you set it up with your own consumables.
Click here to download the user guide.
Click here to download the user guide.
Library Construction: Core offers the following library construction as standard services
Submission Instruction: Please provide >1 µg of total RNA with concentration between 50–200 ng/µl, RIN > 8
Submission Instruction: Please provide >1ug of total RNA with concentration between 50-200ng/ul, RIN>8.
Submission Instruction: Please provide 10pg-1ng of total RNA with concentration < 1ng/uL, RIN>8.
Submission Instruction: Please provide >100ng of DNA with concentration 5-50 ng/ul with A260/280=1.8-2.0, A260/230=2.0-2.2
Submission Instruction: Please provide > 5 ng of DNA with concentration > 0.2 ng/uL with A260/280=1.8-2.0, A260/230=2.0-2.2
Submission Instruction: Please provide >500ng of total RNA with concentration between 50–200 ng/µl, RIN > 8
Submission Instruction: Please provide >500ng of total RNA with concentration between 50–200 ng/µl, RIN > 8
Sequencing: Core uses the following sequencers to address different coverages with different costs and turnaround time.
| Flow Cell Type | Total Lanes (per flow cell) | Output (whole flowcell) | Kits options (per flow cell) |
|---|---|---|---|
| 1.5B | 2 | 1.5B |
100 cycles (max. 138 cycles) 200 cycles (max. 238 cycles) 300 cycles (max. 238 cycles) |
| 10B | 8 | 10B |
100 cycles (max. 138 cycles) 200 cycles (max. 238 cycles) 300 cycles (max. 238 cycles) |
| 25B | 8 | 25B | 300 cycles (max. 338 cycles) |
| Flow Cell Type | Total Lanes (per flow cell) | Output (whole flowcell) | Kits options (per flow cell) |
|---|---|---|---|
| P1 | 1 | 100M |
100 cycles kit (max. 138 cycles) 300 cycles kit (max. 338 cycles) 600 cycles kit (max. 638 cycles) |
| P2 | 1 | 400M |
100 cycles kit (max.138 cycles) 200 cycles kit (max.238 cycles) 300 cycles kit (max. 338 cycles) |
| P3 | 1 | 1.1B |
50 cycles kit (max. 88 cycles) 100 cycles kit (max.138 cycles) 200 cycles kit (max.238 cycles) 300 cycles kit (max. 327 cycles) |
| Flow Cell Type | Total Lanes (per flow cell) | Output (whole flowcell) | Kits options (per flow cell) |
|---|---|---|---|
| HighOutput | 1 | 22–25 M |
75 cycles kit (max. 92 cycles) 150 cycles kit (max.168 cycles) 300 cycles kit (max.318 cycles) |
| MidOutput | 1 | 7–8 M | 300 cycles kit (max.318 cycles) |
| Flow Cell Type | Total Lanes (per flow cell) | Output (whole flowcell) | Kits options (per flow cell) |
|---|---|---|---|
| High Output | 2 | 1B | 150 cycles kit (max. 184 cycles) |
| 300 cycles kit (max. 334 cycles) |
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| 300M | 600 cycles kit (max. 634 cycles) |
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| Medium Output | 2 | 500M | 150 cycles kit max. 184 cycles) |
| 300 cycles kit (max. 334 cycles) |
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| 100M | 600 cycles kit (max. 634 cycles) |
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| Low Output | 2 | 250M | 150 cycles kit (max. 184 cycles) |
| 100M | 600 cycles kit (max. 634 cycles) |
Library Construction: Core offers the following library construction as standard services:
Submission Instruction:
| Input | Total Input | Concentration | QC | Comments |
|---|---|---|---|---|
| Total RNA | > 300ng | 50-200 ng/µl |
A260/280=1.8-2.0 A260/230=2.0-2.2 RIN > 7 |
Resuspend in RNase-free water |
| cDNA | > 1 µg | cDNA Profile (Tapestation or Bioanalyzer) | Resuspend in EB or low EDTA TE |
Submission Instruction:
| Input | Total Input | Concentration | QC | Comments |
|---|---|---|---|---|
| HMW gDNA | > 2µg | 50-100 ng/µl |
A260/280=1.8-2.0 A260/230=2.0-2.2 |
Resuspend in EB or low EDTA TE. **DNA should be at high quality and sufficient quantity since there is no amplification step. |
Submission Instruction:
| Input | Total Input | Concentration | QC | Comments |
|---|---|---|---|---|
| HMW gDNA | > 2µg | 50-100 ng/µl |
A260/280=1.8-2.0 A260/230=2.0-2.2 |
EB, low EDTA TE (pH8) |
Sequencing: We use the lasted technology Revio for fast turnaround time and getting the reliable data.
| Library | Run time* | Q30+ bases | HiFi yield per SMRT Cell | Methylation |
|---|---|---|---|---|
| 1–5 kb | 12 hours | 95% | 6–8 million reads | |
| 5–10 kb | 35–70 Gb | |||
| 10–15 kb | 24 hours | 90% | 70–100 Gb | |
| 15–20 kb | 100–120 Gb | |||
| 20–25 kb | 30 hours | 85% | 100–120 Gb | 5mC at CpG sites and 6mA† for native DNA |
| Library | Application | Per Revio SMRT Cell | Expected coverage |
|---|---|---|---|
| 1–5 kb | Amplicon sequencing | >1,000 | 50× |
| 5–10 kb | Microbial genome | 384 | 30× |
| 5–10 kb | PureTarget repeat expansion panel | 48 | 200× |
| 15–20 kb | Human genome | 2 | 20× |
| 15–20 kb | Human methylation profiling | 8 | 5× |
| 15–20 kb | Transcriptome with Kinnex™ full-length RNA kit | 6 | 10 million reads |