Sample Requirements

Customer provides fluorescently labeled PCR products/ fragments as sample. Refer our requirements tabulated below:

Service Type

Sample Volume

Remarks

a. FA with Spectral Optimization

15uL of undiluted sample

1st BASE to determine the dilution factor (DF) prior electrophoresis.

b. FA without Spectral Optimization [Customer to determine final DF]

Customer may determine the dilution factor to 1st BASE to waive the spectral optimization fees. Our total injection volume in Genetic Analyser is 10uL (which included Size Standard and Formamide). If 1uL of undiluted sample is acquired for Fragment Analysis, the final dilution factor becomes 10. If the sample needs to dilute 4x at 1:3 (PCR template: H2O), the final dilution factor to us thus becomes 40. Please use this calculation to indicate the final dilution factor to us.

c. FA without Spectral Optimization [Customer pre-dilute samples and 1st BASE loads 2uL]

15uL of diluted sample, without addition of any size standard or formamide

More savings if customers choose to directly dilute the samples on their own. With this, 1st BASE takes 2uL of the customer self-diluted DNA for electrophoresis. Recommended dilution is typically between 1uL:3uL – 1uL:90uL (PCR template: dH20). The total injection volume in Genetic Analyser remains as 10uL (which already included 2uL of your samples).

d. FA Ready-to-Load (Customer perform full sample preparation to 1st BASE’s requirements)

15uL of diluted sample, mixed with size standard and formamide

Customers to mix appropriate amounts of PCR template, size standard, and Hi-Di Formamide in 15uL of total solution (minumum volume of 10uL is required). 1st BASE loads the mixture to Genetic Analyzer directly. We only accept Applied Biosystems (ABI) ROX-500 or LIZ-500 size standards ranging in 50, 75, 100, 139, 150, 160, 200, 250, 300, 340, 350, 400, 450, 490 and 500bp.

Submit the samples using strip capped 96-well plate, 8-PCR strips, or 1.5mL micro tubes. Please wrap with aluminum foil before sending it to us.

Tips on packing your samples in a 96-well plate

  • Please place your samples properly into strip-capped well plate as shown below. Adhesive seals or sealing mats will cause mix-up of wells during transportation.

     
  • To minimize the physical damage, it is best to use out-skirted well plate wrapped with parafilm.
     
  • Please ensure equal volume and seal tightly with 8-PCR strip cap to avoid any potential damages in transit such as evaporation or contamination of samples during shipping.
     
  • Bad sealing may lead cross-contamination and unsatisfactory results! Re-sequencing is additionally charged.