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

Blood / Saliva DNA Extraction

Blood:

  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. Add 0.5 to 2.5µL of blood sample (fresh blood, EDTA added blood , dried blood, blood clots) to the tube.
  4. Mix well by tapping the vial with your index finger and make sure the blood mixed completely. The solution turns clear light red/pink once the blood is mixed properly.
  5. The DNA is now extracted and is ready for any downstream application.
  6. Please note that the blood sample can be collected by any method, finger prick or from peripheral veins.
  7. In case of dried blood or blood clot, quantity equivalent to 2.5 µL should be added.
  8. DNA yield may be lower in lysed blood compared to fresh or properly stored blood samples.

Video description

Saliva:

  1. Incubate the bottle of AmpReady® at 37 oC and mix well before each use.
  2. Transfer 600µL of reagent to a fresh 1.5 ml microcentrifuge tube.
  3. Wash mouth with fresh water thoroughly before collecting saliva sample.
  4. Stimulate fresh saliva secretion by squeezing buccal mucosa (most recommended step).
  5. Collect saliva in a fresh tube and try to avoid mucus in the collected sample.
  6. Transfer 1 to 10 µL  to the tube containing AmpReady® reagent and mix well by tapping the vial with your index finger.
  7. The DNA is now extracted and is ready for any downstream application.

Buccal Mucosa:

  1. Incubate the bottle of AmpReady® at 37 oC and mix well before each use.
  2. Transfer 600µL of reagent to a fresh 1.5 ml microcentrifuge tube.
  3. Take a cotton/nylon swab and rub it against the buccal mucosa for 30 seconds.
  4. Cut the tip of the swab and place it in the tube of AmpReady® and mix well by tapping the vial with your index finger.
  5. The DNA is now extracted and is ready for any downstream application.
  6. Please note that the swab should have less absorbent material to avoid complete absorption of solution in AmpReady® tube.

Forensic Samples:

  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. Add the sample (blood stained cloth, dried blood, nails, hair follicle, etc) directly to the tube of AmpReady® and mix well by tapping the vial with your index finger or using micropipette.
  4. Solid samples like nails can be cut into small pieces before adding to the vial.
  5. The DNA is now extracted and is ready for any downstream application

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