MERmaid^® SPIN Kit Protocol
Revision No. 1105-999-5F03W

The MERmaid SPIN Kit is the superior method for purifying small MW DNA away from gel material, nucleotides, salts, proteins, or other undesirable components of a mixture. Single-stranded or double-stranded DNA oligomers or PCR products of 10-200 nucleotides in length are readily purified from agarose and polyacrylamide gels or from solution, without alcohol precipitations or hazardous organic extractions. The new SPIN procedures purify DNA more rapidly than the non-spin version of the MERmaid Kit and eliminate matrix carry-over which can interfere with subsequent enzyme reactions. The procedure is used for simultaneous desalting, buffer exchange, and concentration of DNA. Non-radioactive and radioactive-labeled probes, linkers, and primers are cleaned using the Label Block reagent. When purifying DNA from agarose, especially if <50 bp in length, it is recommended that all MERmaid SPIN Kit reagents be used for the best recovery of DNA. Substituting other agaroses or electrophoresis buffers may result in less than optimal yields in these instances. Purification of DNA from gels containing TBE BUFFERS is often variable and is not recommended for optimum results, although gel slices from TBE gels can be soaked in non-borate containing buffers to dilute the TBE prior to using the MERmaid Kit with satisfactory results.

1. Add GLASSFOG SPIN Bind to MERmaid SPIN Filter; add DNA solution; mix and spin to empty filter.
2. Wash twice with MERmaid SPIN Ethanol Wash*.
3. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.

1. Add GLASSFOG SPIN Bind to MERmaid SPIN Filter; add gel slice; heat to melt gel; mix and spin to empty filter.
2. Wash with MERmaid SPIN High Salt Wash.
3. Wash twice with MERmaid SPIN Ethanol Wash*.
4. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.

1. Add GLASSFOG SPIN Bind to MERmaid SPIN Filter; add crushed polyacrylamide gel slice; heat; mix, and spin to empty filter.
2. Wash twice with MERmaid SPIN Ethanol Wash*.
3. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.

1. Add Label Block to GLASSFOG SPIN Bind before adding to SPIN Filter.
2. Follow the appropriate protocol above for the specific application.

Time Consideration: 10-15 minutes

1. Add 400 µl of GLASSFOG SPIN Bind to MERmaid SPIN Filter; add DNA solution; mix and spin to empty filter.
   1. Shake GLASSFOG SPIN Bind Solution to resuspend GLASSFOG matrix and add 400 µl to a SPIN Filter.
   2. Add up to 10 µg of DNA in up to 300 µl of solution.
   3. Incubate 5 minutes at room temperature with agitation (shaker, rotator, or vortex at less than half maximal speed).
   4. Centrifuge at full speed for approximately 30 seconds to transfer liquid to catch tube.
2. Wash twice with MERmaid Spin Ethanol Wash.
   1. Add 500 µl of MERmaid SPIN Ethanol Wash to the SPIN Filter (add ethanol before first use of MERmaid SPIN Ethanol Wash).
   2. Spin for 30 seconds or until SPIN Filter is emptied of solution. Repeat wash.
   3. Empty catch tube and spin for 1 minute to "dry" pellet.
3. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.
   1. Add 10 to 25 µl of MERmaid Elution Solution to SPIN Filter and resuspend GLASSFOG by flicking the tube.
   2. Spin for 30 seconds to transfer eluted DNA to catch tube.
   
   
   

   A second elution can increase yield 10-15%

   
   
   
   3. Discard SPIN Filter and cap the tube. DNA in solution is ready to use without further manipulation.

Time Consideration: 15 Minutes

1. Add 400 µl of GLASSFOG SPIN Bind to MERmaid SPIN Filter; add gel slice,;heat to melt gel; mix and spin to empty filter.
   1. Shake GLASSFOG¨ SPIN Bind Solution to resuspend GLASSFOG¨ matrix and add 400 µl to SPIN Filter.
   2. Add up to 300 mg of agarose gel slice and heat in a water bath for 5 minutes at 55°C to melt gel.
   3. Incubate 5 minutes at room temperature with agitation (shaker, rotator, or vortex at less than half maximal speed).
   4. Centrifuge at full speed for approximately 30 seconds to transfer liquid to catch tube.
2. Wash with MERmaid SPIN High Salt Wash.
   1. Add 500 µl of MERmaid SPIN High Salt Wash to the SPIN Filter.
   2. Spin for 30 seconds or until SPIN Filter is emptied of wash. Repeat wash if working with 3% or greater agarose concentration.
3. Wash twice with MERmaid SPIN Ethanol Wash.
   1. Add 500 µl of MERmaid SPIN Ethanol Wash to the SPIN Filter (add ethanol before first use of MERmaid SPIN Ethanol Wash).
   2. Spin for 30 seconds or until SPIN Filter is emptied of wash. Repeat wash.
   3. Empty catch tube and spin for 1 minute to "dry" pellet.
4. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.
   1. Add 10 to 25 µl of MERmaid Elution Solution to SPIN Filter and resuspend GLASSFOG by flicking the tube.
   2. Spin for 30 seconds to transfer eluted DNA to catch tube.
   
   
   

   A second elution can increase yield by 10-15%

   
   
   
   3. Discard SPIN Filter and cap the tube. DNA in solution is ready to use with- out further manipulation.

Time Consideration: 15-30 Minutes

1. Add 400 µl of GLASSFOG SPIN Bind to MERmaid SPIN Filter; add crushed polyacrylamide gel slice; heat; mix, and spin to empty filter.
   1. Crush polyacrylamide gel slice with a spatula on a piece of Parafilm or spin in a microcentrifuge tube that has a pinhole in the bottom (make hole with hot needle).
   2. Transfer gel pieces to a SPIN Filter containing 400 µl of GLASSFOG SPIN Bind Solution.
   3. Incubate 5 minutes at room temperature with agitation (shaker, rotator, or vortex at less than half maximal speed).
   4. Heat tube at 55°C for 5-15 (longer times, greater recovery) minutes to allow DNA to diffuse out of gel pieces.
   5. Spin at high speed in microcentrifuge to transfer liquid to catch tube.
2. Wash twice with MERmaid SPIN Ethanol Wash.
   1. Add 500 µl of MERmaid SPIN Ethanol Wash to the SPIN Filter (add ethanol before first use of MERmaid SPIN Ethanol Wash).
   2. Spin for 30 seconds or until SPIN Filter is emptied of wash. Repeat wash.
   3. Empty catch tube and spin for 1 minute to "dry" pellet.
3. Transfer SPIN Filter to an Elution Catch Tube and elute DNA with MERmaid Elution Solution.
   1. Add 10 to 25 µl of MERmaid Elution Solution to SPIN Filter and resuspend GLASSFOG by flicking the tube.
   2. Spin for 30 seconds to transfer eluted DNA to catch tube.




A second elution can increase yield by 10-15%




4. Discard SPIN Filter and cap the tube. DNA in solution is ready to use without further manipulation.

Time Consideration: 15-30 Minutes

When working with isotope or non-isotope labeled DNA, add 20 µl of Label Block to 400 µl of GLASSFOG SPIN Bind before adding to spin filter. This will assist in removal of DNA from the GLASSFOG Matrix. Once the Label Block has been added to the GLASSFOG SPIN Bind, follow the appropriate protocol above for the specific application.