Sanger DNA Sequencing
The most flexible Sanger Sequencing Service! Offering skilled analysis of your sequencing results and expert troubleshooting to optimize more complex samples (e.g., hairpin, GCrich templates). Additional services such as template amplification and PCR purification are also offered.
+ D101: Standard DNA Sequencing
Sanger DNA Sequencing (up to 1000 bp) – Purified Samples
Turnaround Time: 24 hours
Service Description:
BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Purified plasmid DNA or PCR product, in pure H2O; Concentration ≥ 50 ng/µl; OD260/OD280 = 1.8-1.95; 10 µl of sample per sequencing reaction
For ≥15 kb plasmid, 10 µl of sample per sequencing reaction with a concentration ≥100 ng/µl DNA is required.
For purified PCR DNA samples, the concentration required is based on 2.5 ng/µl per 100 bp (for 1 kb PCR sample, 10 µl of 2.5 ng/µl PCR DNA sample is required)
Please submit sequencing samples in 8-strip PCR tubes
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D102: GC-rich DNA Sequencing
GC-rich Sanger DNA Sequencing (up to 1000 bp) – Purified Samples
Turnaround Time: 24 hours
Service Description:
BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Purified plasmid DNA or PCR product, in pure H2O; Concentration ≥ 50 ng/µl; OD260/OD280 = 1.8-1.95; 10 µl of sample per sequencing reaction
For ≥15 kb plasmid, 10 µl of sample per sequencing reaction with a concentration ≥100 ng/µl DNA is required.
For purified PCR DNA samples, the concentration required is based on 2.5 ng/µl per 100 bp (for 1 kb PCR sample, 10 µl of 2.5 ng/µl PCR DNA sample is required)
Please submit sequencing samples in 8-strip PCR tubes
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D103: PCR Direct Colony Sequencing
Sanger DNA Sequencing (up to 1000 bp) – Bacterial Colony Samples
Turnaround Time: 24 hours
Service Description:
Colony lysis, PCR, BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Single colony on LB Agar plate or single colony bacterial culture in LB medium.
Forward and reverse sequencing primers are required (5 mM, 4µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D104: PCR Cleanup & DNA Sequencing
Sanger DNA Sequencing (up to 1000 bp) – Unpurified PCR Samples
Turnaround Time: 24 hours
Service Description:
PCR product purification, BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
≥ 10 µl of PCR product with a specific target PCR DNA band confirmed by agarose gel electrophoresis
Please submit sequencing samples in 8-strip PCR tubes
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D105: PCR Cleanup & GC-rich Sanger DNA Sequencing
GC-rich Sanger DNA Sequencing (up to 1000 bp) – Unpurified Samples
Turnaround Time: 24 hours
Service Description:
PCR product purification, BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
≥ 10 µl of PCR product with a specific target PCR DNA band confirmed by agarose gel electrophoresis
Please submit sequencing samples in 8-strip PCR tubes
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D106: Plasmid Miniprep & DNA Sequencing
Sanger DNA Sequencing (up to 1000 bp) – Unpurified Plasmid Samples
Turnaround Time: 48 hours
Service Description:
Plasmid Miniprep, BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Single colonies on LB Agar plate or single colony bacterial culture in LB medium
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D107: RCA Direct Colony Sequencing
RCA Sanger DNA Sequencing (up to 1000 bp) – Bacterial Colony Samples
Turnaround Time: 48 hours
Service Description:
RCA, BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Single colonies on LB Agar plate or single colony bacterial culture in LB medium
One Sequencing primer is required (5 mM, 2µl per sequencing reaction)
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
+ D108: Standard DNA Sequencing Tubes
Standard Sanger DNA Sequencing (up to 1000 bp)
Turnaround Time: 24 hours
Service Description:
BigDye™ Terminator v3.1 Cycle Sequencing
Deliverables:
Sequence data in .seq and .ab1 file formats
Suitable Sample Type:
Minimum Quantity = 48
• Identification of proteins from protein gel bands or gel spots
• Up to 2,000 proteins identified from one gel band
• Workflow: Sample preparation, in-gel enzymatic digestion, peptide extraction, LC-MS/MS analysis, data analysis, and results report
• Protein Identification Report
• List of identified proteins
• List of identified proteins with matched peptides sequences
• Raw data files, upon request
• Gel band or spot, no larger than 3mm (l) x 3.5mm (h) x 1.5mm (w) in size (Note: additional charge applied for larger size)
Sample Submission Guidelines
Premix Primer With DNA Sample
Premixed for success
Advantages of premixed samples:
- Track samples easily
- Get results faster
- Enjoy low cost per reaction
- Improve success rate significantly
Mixed in the same tube: Template + Primer
Unpremixed, DNA and Primer In Separate Tubes
Tube 1: Template Tube 2: Primer Primer (5 pmol/μl or 5 μM), 10 μl. Or, use our free primer library.
Primer For Sequencing: Prefer Tm: 50-60C, GC content: 40-60%
E. Coli Colonies
Pick a colony and resuspend in diH2O or Tris Buffer
Two sets of tubes:
Tube 1: Colony Suspension Tube 2: Primer
- Colonies need to grow for at least 16hrs at 37°C to reach good visible size
- Pick a single colony with a sterile tip and resuspend in 30ul sterile diH2O or Tris buffer (10mM, ph8.0), 15 ul will be submitted to us, and the remaining 15 ul will be kept by the clients for future uses
- Prepare 2 separate tubes: 1 contains 15ul colony suspension, 1 contains 5ul primer at 5pmol/ul
How to Label Sample Tubes
Label the samples with your initials followed by numbers
8-strip PCR tube for samples containing template
Example: Samples from John Smith
Label on the cap as well as on the side of the tube
Features & Benefits
- High Quality Result
- Overnight turnaround time
- Expert technical support
- Daily Pick up with various Drop Box locations
- Free Universal Primer
- Free repeats on failed reactions
