Preparation of Samples for DNA sequencing

DNA Sequencing Quick Guide

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

Sample Preparation Requirements for DNA Sequencing Reactions for local customers (Singapore and Malaysia)

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

 

Type of Primer

Required Concentration

Total of Reaction

Required Volume

Sequencing Primer

  • 18 to 25 bases
  • GC content: 40% to 60%
  • Tm = 55°C to 60°C
  • No significant hairpins (>3bp) and homodimerization
  • Desalted Primer (Brand: IDT) or Purified Primer (other Manufacturer)

10µM or 10pmol/µL or 50ng/uL

1

10uL

10

25uL

20

50uL

 

Sample Preparation Requirements 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

 

Type of Primer

Required Concentration

Total of Reaction

Required Volume

Sequencing Primer

  • 18 to 25 bases
  • GC content: 40% to 60%
  • Tm = 55°C to 60°C
  • No significant hairpins (>3bp) and homodimerization
  • Desalted Primer (Brand: IDT) or Purified Primer (other Manufacturer)

10µM or 10pmol/µL or 50ng/uL

1

10uL

10

25uL

20

50uL

Packing of Samples (DNA templates and primers)

  1. Please prepare Sample in microcentrifuge tube (preferably in 0.5mL tubes) or directly on 96-well plate for high-throughput sequencing orders (empty the well no. H12 for our internal control reaction)
  2. Label clearly on the side and top of tube with a permanent marker
  3. Please use simple labels on your tubes (e.g. initials and numbers) and avoid long sample names
  4. Ensure that the lid is tightly closed and seal the tubes with parafilm
  5. 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:

  1. Please prepare Samples in microcentrifuge tube (preferably in 0.5mL 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)
  2. Label clearly on the side and top of tube with a permanent marker
  3. Please use simple labels on your tubes (e.g. initials and numbers) and avoid long sample names
  4. Ensure that the lid is tightly closed and seal the lid with parafilm
  5. Place samples in a plastic bag. Do not stick samples directly onto order forms, please attach order form to the plastic bag instead.
  6. 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.
  7. 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 and Singapore Customers

How to Pack and Seal 96-Well for International Customers