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YOSHITAKA HISHIKAWA ET AL.
1. Find quartz dishes, two forceps and a sheet of Parafilm.
2. Switch on UV lamps at least 15-30 mins before use.
3. Check and adjust position of the stand to give a dose of 5JI
m 2 /sec by using UV-radiometer.
4. Wash a quartz dish with sterile DDW before use.
5. Apply 60 fll of DNA solution (50 flg/ml)( depending upon the
capacity of a quartz dish) in several drops onto the dish and
seal it with a quartz cover.
Note: Be careful not to contaminsate DNA solution and the inside of the quartz dish with DNase and RNase at this step.
6. Place a set of the dish onto a platform of the stand and allow
it to be irradiated for an appropriate time.
Note: 5000 J/m 2 : irradiation 1000 secs with UV lamp (5 J/m2/sec)
10000 J/m2 : irradiation 2000 secs with UV lamp (5 J/m2/sec)
12000 J/m2 : irradiation 2400 secs with UV lamp (5 J/m2/sec)
7. Take out the dish and put it on a sheet of Parafilm.
8. Using forceps, remove the cover and collect the DNA solution.
9. Immediately after use of the dishes, immerse them into sterile DDW. Don't dry up completely.(*)
Note: (*) If the quartz dish is dried up completely before rinsing
or if you feel the dish was stained with DNA or unwanted objects,
skip to the next step.
1 *. Immerse the quartz dish into 2% Adosolve in DDW for 30
mins at RT.
2*. Rinse them three times with DDW for 10 mins each.
3*. Immerse them into 0.2% DEPC for 20 mins at RT.
4*. Rinse them three times with DDW for 10 mins each.
5*. Carry out steps 4 and 5 thoroughly.
10. Rinse the quartz dishes two more times with sterile DDW.
11. Rinse them once with ethanol.
12. Return them to the original plastic dish and allow to dry.
YOSHITAKA HISHIKAWA ET AL.
1. Find quartz dishes, two forceps and a sheet of Parafilm.
2. Switch on UV lamps at least 15-30 mins before use.
3. Check and adjust position of the stand to give a dose of 5JI
m 2 /sec by using UV-radiometer.
4. Wash a quartz dish with sterile DDW before use.
5. Apply 60 fll of DNA solution (50 flg/ml)( depending upon the
capacity of a quartz dish) in several drops onto the dish and
seal it with a quartz cover.
Note: Be careful not to contaminsate DNA solution and the inside of the quartz dish with DNase and RNase at this step.
6. Place a set of the dish onto a platform of the stand and allow
it to be irradiated for an appropriate time.
Note: 5000 J/m 2 : irradiation 1000 secs with UV lamp (5 J/m2/sec)
10000 J/m2 : irradiation 2000 secs with UV lamp (5 J/m2/sec)
12000 J/m2 : irradiation 2400 secs with UV lamp (5 J/m2/sec)
7. Take out the dish and put it on a sheet of Parafilm.
8. Using forceps, remove the cover and collect the DNA solution.
9. Immediately after use of the dishes, immerse them into sterile DDW. Don't dry up completely.(*)
Note: (*) If the quartz dish is dried up completely before rinsing
or if you feel the dish was stained with DNA or unwanted objects,
skip to the next step.
1 *. Immerse the quartz dish into 2% Adosolve in DDW for 30
mins at RT.
2*. Rinse them three times with DDW for 10 mins each.
3*. Immerse them into 0.2% DEPC for 20 mins at RT.
4*. Rinse them three times with DDW for 10 mins each.
5*. Carry out steps 4 and 5 thoroughly.
10. Rinse the quartz dishes two more times with sterile DDW.
11. Rinse them once with ethanol.
12. Return them to the original plastic dish and allow to dry.