Array Advice - Quantitation

View tips and solutions to common problems for sample quantitation.

Tips

  • Quantitate your gDNA using a Nanodrop spectrophotometer or a fluorometer e.g. Qubit.

Nanodrop and Qubit

  • Spectrophotometers can over estimate the amount of DNA. For lower concentrations, use a fluorescent assay in addition to spectrophotometer (e.g. Qubit).
  • Ensure you use the correct blanking buffer with the spectrophotometer. The blanking buffer should be the same as the buffer that the DNA sample is dissolved in.

Solutions to Common Problems

Problem 1 - Inaccurate sample concentration

Cause: Sample concentration should be <400ng/ul. Inaccurate sample concentration can be caused because gDNA at high concentration cannot be quantitated accurately. 

Solution: If the gDNA concentration is > 350 ng/μl, dilute 1:2 with water (or any of the recommended buffers).

 

Problem 2 - Precipitated DNA

Cause: Precipitated DNA can be caused by gDNA not being completely dissolved.

Solution: To resolve make sure that the gDNA is completely in solution by pipetting up and down. If needed, incubate at 37°C for 30 minutes.

 

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