Pyrosequencing service (Pyro@INFORMM)

Pyrosequencing is a type of DNA sequencing based on the “sequencing by synthesis” method. Unlike traditional Sanger sequencing, Pyrosequencing does not require fluorescence dye labelling or gel electrophoresis, while allowing quantitative, real time detection of sequencing events. Pyro@INFORMM is offering a pyrosequencing service for SNP and mutational analysis as well as quantitative analysis of CpG methylation. Please find below the details of the packages offered by us.

SERVICE CHARGES

No. Package Price (RM)
1.

Primer design using “Pyromark Assay Design 2.0” software
Note: Primer design by users

 FOC
2.

Machine handling

Note: i. Consumables and reagent provided by user
        ii. Capacity: Maximum 88 reactions/ run

RM 70.00/day

Up to 3 run/day
3.

Full service
Note: Consumables and reagent provided by INFORMM

 RM 200.00/reaction

SERVICE ORDERING PROCEDURE

SAMPLE PREPARATION GUIDELINES

  • Prepare biotin-labelled PCR fragments (Sequence to be analyzed) according to the assay design
  • The PCR primer that is complementary to the sequencing primer MUST be HPLC-purified and biotin-labelled
  • Use no more than 0.2uM PCR primers with high number of PCR cycle to deplete excess biotin-labelled primers. Excess biotin-labelled primers will decrease assay efficiency
  • The PCR products MUST be specific and clean (i.e. without primer dimers and non-specific products)
  • 2-5 pmoles of PCR product are required per reaction. We strongly recommend quantifying of the PCR products by running a DNA Mass ladder in the same agarose gel

SAMPLE SUBMISSION

  • Prepare 2-5 pmoles of PCR products in a normalized volume between 20-50 ul in a 96-well plate.
  • Load maximum 88 samples per plate in columns 1 to 11 only.
  • Seal the 96-well plate with adhesive film securely.
  • Label the plate clearly with your name and date.
  • Dilute the sequencing primer to 20uM and prepare at least 1.5ul per sample in 1.5 ml microtube.
  • Label the tube clearly with primer name and date.
  • Put the plate and tube(s) in clear plastic bag.

CONTACT PERSON

Mr. Muhammad Fazli Khalid (fazlikhalid@usm.my; +609-7672428)
Miss Basyirah Ghazali (basyirah@usm.my; +609-7672414)

Administration office Tel: +609-7672403