Sanger Sequencing Services Order Preparation and Submission
Please ensure that your samples meet our requirements.
Download Sample Preparation Quick Guide below to aid the speedy preparation of your samples.
| Singapore Customers | Malaysia Customers | International Customers |
| DNA Sequencing Quick Guide | DNA Sequencing Quick Guide | DNA Sequencing Quick Guide |
Single Pass DNA Sequencing Sample Requirements
For Singapore & Malaysia Customers
| Type of DNA Template | Recommended Concentration/ Reaction | Required Volume/ Reaction |
| Purified PCR Product [90-500bp] | 5 ng/µL | 10 uL |
| Purified PCR Product [501-1500bp] | 20 ng/µL | |
| Purified PCR Product [>1500bp] | 30 ng/µL | |
| Purified Plasmid (< 4kb) | 50 ng/µL | |
| Purified Plasmid (4kb to 10kb) | 80 ng/µL | |
| Purified Plasmid (>10kb) | 100 ng/µL | |
| Purified M13/ phagemid ssDNA | 50 ng/µL | |
| Purified Genomic DNA/ BAC | 400 ng/µL | 20 uL |
For International Customers
We recommend that international customers send a larger volume of their samples to overcome loss of samples due to evaporation during the extended transportation to our facilities.
| Type of DNA Template | Recommended Concentration/ Reaction | Required Volume/ Reaction |
| Purified PCR Product [90-500bp] | 5 ng/µL | 15 uL |
| Purified PCR Product [501-1500bp] | 20 ng/µL | |
| Purified PCR Product [>1500bp] | 30 ng/µL | |
| Purified Plasmid (< 4kb) | 50 ng/µL | |
| Purified Plasmid (4kb to 10kb) | 80 ng/µL | |
| Purified Plasmid (>10kb) | 100 ng/µL | |
| Purified M13/ phagemid ssDNA | 50 ng/µL | |
| Purified Genomic DNA/ BAC | 400 ng/µL | 25 uL |
Sequencing+ PLUS Sample Requirements
For Unpurified Samples
| Sample Type | General Guideline |
| Unpurified PCR Products | 20µL in 1.5ml tube/ plate Visible single DNA band on agarose gel with 1ul of volume. |
| Excised Gel Band | <250mg in 1.5mL tube Visible DNA band at the required target size on agarose gel. |
For Plasmid (Singapore Customers)
Click here to enlarge image.
Direct Colony Sequencing Sample Requirements
| Sample Type | Colony Size |
Slant surface agar | Pick a single colony using sterile inoculating loop and streak on the surface of the prepared agar. Incubate the culture at optimum growing temperature for overnight. Ensure that there is visible growth of the bacteria colony to the indicated size in Figure below:
Size of the colony should be approximately 1mm (width) X 6 mm (length). NOTE: Do not stab your culture into the agar. The colony should grow on the surface of the agar for better visibility of the colony size. |
| Straight surface agar
|
M13-Tailed Sequencing Sample Requirements
For Malaysia & International Customers
Sample Type | Purification | Specification | Size and Volume | Disclaimers |
qPCR Products | Only unpurified qPCR/ PCR products are accepted | Ct value < 33 | > 90 bp, minimum 15µL | (i) M13-tailed amplification needs ~ 5ng of DNA amount for each reaction. |
PCR Products | Visible on agarose gel using 2µL | > 90 bp, minimum 15µL |
Primer Walking Sample Requirements
Length | Concentration & Volume/ Project | |
Primer Walking [<= 3kb] | 100 ng/µL, min 50 µL per rxn in dH2O | 1.5kb per week |
Primer Walking [> 3kb] | Whole bacterial colony OR 100 ng/µL, min 50 µL per rxn in dH2O | |
Primer Walking Sample Requirements
Length | Concentration & Volume/ Project | |
Primer Walking [<= 3kb] | 100 ng/µL, min 50 µL per rxn in dH2O | 1.5kb per week |
Primer Walking [> 3kb] | Whole bacterial colony OR 100 ng/µL, min 50 µL per rxn in dH2O | |
Sample Preparation Requirements for DNA Purification & Quantification Services
We recommend that international customers send a larger volume of their samples to overcome loss of samples due to evaporation during the extended transportation to our facilities.
Sample Preparation and Quantification Service | Malaysia & Singapore | International |
Required Concentration & Volume | ||
Quantification by UV Spectrometry (SS1021) | min. 15 µL for each sample | min. 25 µL for each sample |
Quantification by Agarose Gel (SS1020) | min. 15 µL for each sample | min. 25 µL for each sample |
Purification Using PCR Clean-up (SS1012/3) | 100 ng/µL, min 20 µL per reaction in dH2O | 100 ng/µL, min 40 µL per reaction in dH2O |
Purification Using Gel Extraction (SS1014/5) | 100 ng/µL, min 30 µL per reaction in dH2O | 100 ng/µL, min 40 µL per reaction in dH2O |
Ready-to-Load Sequencing with Clean-up (SS1019) | min. 10 µL, please indicate cycle reaction volume. | min. 20 µL, please indicate cycle reaction volume. |
Sequencing Primers Requirements
| Type of Primer | Required Concentration | Total of Reaction | Required Volume |
Sequencing Primer
| 10µM or 10pmol/µL or 50ng/uL | 1 | 10uL |
| 10 | 25uL | ||
| 20 | 50uL |
Packing of Samples (DNA Templates and Sequencing Primers)
- Please prepare Sample in microcentrifuge tube (preferably in 1.5mL tubes) or directly on 96-well plate for high-throughput sequencing orders (empty the well no. H12 for our internal control reaction)
- Label clearly on the side and top of tube with a permanent marker
- Please use simple labels on your tubes (e.g. initials and numbers) and avoid long sample names
- Ensure that the lid is tightly closed and seal the tubes with parafilm
- Place samples in a plastic bag and attach the completed DNA Sequencing order form
Guidelines for packaging for safe transportation of DNA sequencing samples from Outstationed / International Customers:
- Please prepare Samples in microcentrifuge tube (preferably in 1.5mL tubes) or directly on 96-well plate sealed with strip-caps for high-throughput sequencing orders (empty the well no. H12 for our internal control reaction)
- Label clearly on the side and top of tube with a permanent marker
- Please use simple labels on your tubes (e.g. initials and numbers) and avoid long sample names
- Ensure that the lid is tightly closed and seal the lid with parafilm
- Place samples in a plastic bag. Do not stick samples directly onto order forms, please attach order form to the plastic bag instead.
- Wrap the plastic bag with a bubble bag and send in a padded envelope by overnight delivery either through Courier. DNA samples can be sent at room temperature.
- Notify 1st BASE via email and provide tracking number if any.
When in doubt, do not hesitate to contact us for any clarifications.
How to Pack and Seal 96-Well for Malaysia & Singapore Customers
How to Pack and Seal 96-Well for International Customers