[PRECELLYS] DNA Extraction from Chlorella sorokiniana (Algae) - Bertin Technologies

[PRECELLYS] DNA Extraction from Chlorella sorokiniana (Algae)

Context

Precellys®24 has been used by Promega internal application lab to extract gDNA from Chlorella sorokiniana, before sequencing (NGS). The efficiency of extraction with Precellys®24 has been compared to Proteinase K method based on heating treatment. DNA purification was performed using Promega Maxwell 16 equipment with the LEV Plant DNA kit.

Materials

  • Precellys®24 (Bertin Technologies)
  • Lysing Kit: VK05_2mL (KT03961-1-004.2)
  • Sample: 50 mL of Chlorella sorokinianasuspension (overnight culture) / Several initial volumes of culture (2, 4, 5, 6 and 10 mL)
  • Promega Maxwell 16 LEV Plant DNA Kit (AS1420)
  • Promega Maxwell 16 Instrument (AS3000)
  • NanoDrop 2000

Protocol

  • Centrifugation of each Chlorella sorokinianasuspension (2, 4, 5, 6 and 10 mL)
  • Resuspension of the pellet in 500µL of TLA Buffer (Promega)
  • Precellys® protocol: 6500 rpm, 2 cycles of 20s with 5s of break
  • Heating parameters: 60 min / 70?
  • DNA purification with Promega Maxwell 16 LEV Plant DNA Kit and Maxwell 16 Instrument
  • Quantification of gDNA by NanoDrop
  • Electrophoresis

Results

DNA extraction yields obtained with Precellys®24 or with the Proteinase K method have been evaluated by NanoDrop quantification.

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High  extraction  yields  are  obtained  with  Precellys ® 24 compared to the Proteinase K method. The ratios 260/280 and 260/230 are also very good with Precellys ® 24.

Thanks to the Precellys ® 24, high quality DNA  is extracted from Chlorella sorokiniana in high quantity.

Customer

Promega

Conclusion

Compared to traditional sample preparation methods like proteinase K lysis, Precellys®24 considerably increases the yield of extraction and also the quality of extracted DNA from Chlorella sorokiniana suspension.

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