Cleveland Family Study

5.3.2 Processing Procedure

1. Turn on hot plate containing beaker of distilled water and heat water to 95ºC.
2. Place microcentrifuge tubes in small circular holder and place into water bath for approximately ten minutes. Holder should float on the surface of the water.
3. After ten minutes, remove the holder from the water bath.
4. Mix or shake the tubes briefly.
5. Remove brush using a pair of tweezers and discard. To avoid contamination between samples, wipe the tip of the tweezers with a gauze square before placing the tweezers into another microcentrifuge tube. Samples may be stored at this stage at room temperature for at least 7 days.
6. Neutralize solution with 50uL (about three drops) of 1.0 M Tris-Cl buffer, pH 8.0.
7. Mix well.
8. Remove the tape on the sides of the tubes with the individual’s name.
9. Write the Person ID number on the side of the tube with a permanent marker.
10. Store at 4ºC in freezer.
11. Log the specimen in the “BUCCAL” book. This is a crucial step!
12. Weekly take buccal sample to the –70ºC freezer located on the 6th floor of RB&C in Paul Hartman’s office, room 692. The Family Sleep Apnea samples are stored on the top shelf.
13. Locate the last box used and put the buccal samples in their new location.
14. Document the box number, and location within the box. For example, box 19, location A1 (A= the row and 1 = the column).
15. Bring this information back to the office to be entered into the excel database.

• The excel data base can be found at n:\common\family\family blood data\allbuccul.xls.