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Sample Preparation & Requirements

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Preparation of DNA samples for genotyping and methylation analysis

Please consult platform specialist before sample preparation.

For Genotyping
Isolate DNA using high quality purification reagents/kits and dissolve/elute DNA in TE buffer (10 mM Tris, 1 mM EDTA), pH 8.0.

DNA Sample Quality and Quantity requirements:

  • Double-stranded, non-degraded high quality DNA*
  • Fragment sizes of at least 2 Kb
  • A260/280 = 1.8-2.2
  • Concentration MUST be normalized to 50 ng/uL**
  • Minimum 15 uL**
  • Dissolved in TE buffer(10 mM Tris, 1 mM EDTA), pH 8.0

* Whole Genome Amplified DNA is not recommended.
** Accurate quantification is critical. 
Users are highly recommended to determine sample concentration by fluorometric assay, e.g. use picogreen assay or Qubit (available at CPOS Genomics Core).  Alternative indirect methods such as UV spectrometry/NanoDrop can incorrectly report the concentration of dsDNA in your sample when single-stranded DNA, oligonucleotides, RNA, and/or proteins carried over from DNA extraction are present in the samples.  If these latter methods are used, it is safer to submit 20uL of DNA at 150 ng/uL for Qubit measurement.

Normalized samples should be stored at -80oC until sample submission.


For Methylation Analysis

Isolate DNA using high quality purification reagents/kits and dissolve/elute DNA in TE buffer (10 mM Tris, 1 mM EDTA), pH 8.0.

DNA Sample Quality and Quantity requirements:

  • Double-stranded, non-degraded high quality DNA*
  • Fragment sizes of at least 2 Kb
  • A260/280 = 1.8-2.2
  • Dissolved in TE buffer(10 mM Tris, 1 mM EDTA), pH 8.0

* Whole Genome Amplified DNA is not recommended.

For methylation analysis, sample quantity requirement depends on sample size. Please consult.


For both genotyping and methylation analysis, please examine DNA quality also on 0.8% agarose gel along with proper size marker (with good resolution). 
Retain a softcopy of gel photo to be emailed to platform specialist.
Below is an acceptable example of gel photo:
Sample Gel Photo

 

Contact

  Ms Jade Leung
  Ms Rachel Li
  Ms Lilian Cheng
  Mr Carlos Choi
  Ms Schwan Lui
  Ms Levina Lam

  Telephone: 2831 5467 / 2831 5477 / 2831 5478
  Email: iscan.cpos@hku.hk