AlanFung:Protocol/Zymo EZ DNA Methylation Lighting Kit: Difference between revisions

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>Alan6017518
(Created page with "* Add 130ul lighting conversion reagent to 20ul of sample, compensate with water if less than 20ul. * Vortex and centrifuge. * (98C 8min > 54C 60min > 10C hold) * Add 600ul of...")
 
>Alan6017518
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* Centrifuge at full speed for 30s. Discard flow through.
* Centrifuge at full speed for 30s. Discard flow through.
* Add 100 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30s.
* Add 100 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30s.
* Add 200 μl of L-Desulphonation Buffer to the column and let stand at RT for 120 mins. After the incubation, centrifuge at full speed for 30 seconds.
* Add 200 μl of L-Desulphonation Buffer to the column and let stand at RT for 20 mins. After the incubation, centrifuge at full speed for 30 seconds.
* Add 200 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30 seconds.  
* Add 200 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30 seconds.  
* Repeat this wash step.
* Repeat this wash step.
* Place the column into a 1.5 ml microcentrifuge tube and add 10 μl of M-Elution Buffer directly to the column matrix. Centrifuge for 30 seconds at full speed to elute the DNA.
* Place the column into a 1.5 ml microcentrifuge tube and add 10 μl of M-Elution Buffer directly to the column matrix. Centrifuge for 30 seconds at full speed to elute the DNA.
* The DNA is ready for immediate analysis or can be stored at or below -20°C for later use. -80C for long term storage.
* The DNA is ready for immediate analysis or can be stored at or below -20°C for later use. -80C for long term storage.

Latest revision as of 20:29, 15 July 2015

  • Add 130ul lighting conversion reagent to 20ul of sample, compensate with water if less than 20ul.
  • Vortex and centrifuge.
  • (98C 8min > 54C 60min > 10C hold)
  • Add 600ul of M-Binding Buffer to column with collection tube.
  • Load the sample to column. Close the cap and mix by inverting the column several times.
  • Centrifuge at full speed for 30s. Discard flow through.
  • Add 100 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30s.
  • Add 200 μl of L-Desulphonation Buffer to the column and let stand at RT for 20 mins. After the incubation, centrifuge at full speed for 30 seconds.
  • Add 200 μl of M-Wash Buffer to the column. Centrifuge at full speed for 30 seconds.
  • Repeat this wash step.
  • Place the column into a 1.5 ml microcentrifuge tube and add 10 μl of M-Elution Buffer directly to the column matrix. Centrifuge for 30 seconds at full speed to elute the DNA.
  • The DNA is ready for immediate analysis or can be stored at or below -20°C for later use. -80C for long term storage.