Quick Protocol for Monarch® DNA Gel Extraction Kit ( NEB # T1020 )
Excise the DNA fragment from the agarose gel , taking care to trim excess agarose .
Transfer to a 1.5 ml microfuge tube and weigh the gel slice .
Minimize exposure to UV light .
Add 4 volumes of Gel Dissolving Buffer to the gel slice ( e . g . , 400 μl buffer per 100 μl or 100 mg agarose ) .
Incubate the sample between 37–55°C ( typically 50°C ) , until the gel slice is completely dissolved ( generally 5–10 minutes ) .
The time that takes a gel slice to melt depends on the size of the slice , the temperature used in the incubation as well as the percent agarose used in the gel .
The time recommended above should be used just as a guideline .
Insert column into collection tube and load sample onto the column .
Spin for 1 minute at 16,000 x g , then discard flow - through .
Re - insert column into collection tube .
Add 200 μl DNA Wash Buffer ( with ethanol added ) and spin for 1 minute at 16,000 x g .
Discarding flow - through is optional .
Repeat Step 5 ( Step 5 : Re - insert column into collection tube .
Add 200 μl DNA Wash Buffer and spin for 1 minute at 16,000 x g .
Discarding flow - through is optional ) .
Transfer column to a clean 1.5 ml microfuge tube .
Use care to ensure that the tip of the column does not come into contact with the flow - through .
If in doubt , re - spin for 1 minute .
Add ≥ 6 μl of DNA Elution Buffer to the center of the matrix .
Wait for 1 minute , and spin for 1 minute at 16,000 x g to elute the DNA .
