Thank you for choosing the world's unique DNA extraction technology. Please read the protocol carefully.

Plant DNA Extraction

DATA SHEET

Leaves:

  1. Incubate the bottle of AmpReady® at 37 oC and mix well before each use.
  2. Transfer 200µL of reagent to a fresh 1.5 ml microcentrifuge tube.
  3. Collect a fresh leaf from the plant of your interest and clean the surface with 70% Ethanol. Make sure the leaf is clean from dust particles.
  4. Excise 2mg to 50mg of leaf (Optimal - 5mg) and drop carefully into the tube of AmpReady®.
  5. Homogenize the leaf piece mildly with micropipette tip/micro pestle and mix well  by tapping the tube using your index finger.
  6. The DNA is now extracted and is ready for any downstream application.

Seeds:

  1. Incubate the bottle of AmpReady® at 37 oC and mix well before each use.
  2. Transfer 200µL of reagent to a fresh 1.5 ml microcentrifuge tube.
  3. Take one or two seeds (fresh or dried) and clean with 70% Ethanol.
  4. Collect 5mg to 50mg of seed embryo (Optimal - 10mg) by slicing/scratching (with or without seed coat).
  5. Transfer the pieces to the tube of AmpReady® and homogenize mildly with micropipette tip, mix well  by tapping the tube using your index finger.
  6. The DNA is now extracted and is ready for any downstream application.

Note: The same protocol can be utilized for extracting DNA from radicle of sprouted seeds or root ends.

Quantification of Extracted DNA:

DNA extracted with AmpReady® can be quantified using fluorometric assay (eg. Qubit). Semi-quantitative analysis or visualization of extracted DNA can be performed by loading 10µL of extracted sample directly in 0.8 to 1% agarose gel, followed by electrophoresis and visualization under UV light.

Performing PCR/qPCR or other downstream applications:

Any downstream applications like PCR, qPCR, etc., can be performed directly with the extracted DNA without any additional processing steps. Add 0.5 to 5µL of extracted sample as DNA template to 50µL of your reaction mixture. The template volume can be adjusted according to your standardized reaction conditions.

Tips for your PCR or other applications:

Problem Reason Solution
No / Low intense band / amplification High/Low sample Volume Use recommended quantity of  sample
High/Low template volume Optimize the required template volume (0.5 to 5 µL)
Primer not working Check your primers & annealing temperature
Smear in agarose gel Degradation of primers Check your primers. Running a no template control is recommended
Degradation of DNA template Follow the recommended storage conditions

Storage Conditions:

  • AmpReady® vials can be stored at room temperature up to six months.
  • The extracted DNA samples can be stored as such at 4oC up to 30 days and at -20oC for long term storage.

In case of any queries, please contact:  info@30mgenomics.com