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DNA Cheek Cell Lab

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									                                  NYU Steinhardt School of Culture, Education, and Human Development: Science Lesson Plan Submission




     Science Lesson Plan Submission
     Profile Report
     Date Published: 12/19/2008




     Page One

              1. Lesson Plan Title:

                   DNA cheek cell lab with the use of centrifuge

              2. Subject Area:

                   Living Environment

              3. Topic(s):

                   DNA

              4. Suggested Grade Level:

                   10th

              5. Standards Used and Source:

                   Source:: NYS Living Environment Core Curriculum

                   Standard(s) Used:: performance indicator 2.1 / appropriate lab procedures

              6. Size of class:

                   24-28

              7. Please List Supplies Used and the number of each (1 per line):

                   incubator for class size of test tubes
                   5 centrifuges (or as many needed to hold 1 test tube per student
                   large bottle of gatorade
                   2 test tubes per student
                   about 50mL of detergent solution (to act as a lysis buffer)
                   about 20mL "NaCl and Protease" solution (aka Proteinase K) can use meat tenderizer
                   (the lysis buffer and proteinase acts to break cell membranes)
                   about 50mL of 5M NaCl
                   a couple large bottles of cold 95% isopropyl alcohol or ethanol
                   class set of clean pipettes
                   dixie cups for gatorade (good to have 2 per student)

              8. Do any of these supplies need to be ordered in advance?

                   Yes

              9. Which ones? (1 per line)

                   centrifuge
                   incubator




Page 1 of 3                                                                            Report generated by SurveyGizmo on Dec-19 '08
                                 NYU Steinhardt School of Culture, Education, and Human Development: Science Lesson Plan Submission



              10. Post the lesson text or link here, or upload the lesson or extra materials in the next
              section:

                   2 Day Lab: Obtaining DNA from cheek cells with the use of a centrifuge

                   Protocol:

                   Obtain a test tube from the beaker near the centrifuge near you. Record
                   the number and letter of this tube in your notebook.

                   Obtain a sample of "buccal rinse solution" (it may be Gatorade) in a small
                   cup. It should contain about 15 ml of solution.

                   Vigorously swish the small volume (about 15 mL) of Gatorade in your mouth for 30
                   seconds. Chew on your cheeks while swishing so you get as many cells as possible.
                   Remember: more cells = more DNA!!!

                   Spit the Gatorade back into your cup, then pour about 3cm (about 1 inch or about the length
                   of the end of your thumb) into one of the tubes provided
                   for you (from the centrifuge area) Record your tube number and letter in
                   your notebook.

                   Find a partner using the same centrifuge and make sure your tubes each
                   have the same volume. Add more Gatorade if necessary to make the volumes equal. Place
                   your tubes opposite each other in the appropriate centrifuge. Spin the tubes on high for 3-5
                   minutes to collect cells at the bottom of the tube.

                   Pour the supernatant (the liquid above the pellet of cells that has precipitated on the bottom)
                   into a waste container.

                   Repeat steps #1-4. Pour the Gatorade/Spit from your second mouthwash into the tube with
                   the cells from your first mouthwash. This time do not discard supernatant

                   Using the droppers provided, add about 2.0 mL Detergent solution (Lysis
                   Buffer) to the cell pellet. Mix by carefully flicking the tube.

                   Add 0.50 ml "NaCl and Protease" solution (aka Proteinase K). Flick the
                   tube to break up the cell pellet.

                   Incubate the cells at 65-70 C overnight.


                   DAY 2:
                   Before proceeding, the solution should be clear and the cells should not
                   be visible.

                   Add 1.0 mL of 5M NaCl.

                   Mix well, then centrifuge for 10 minutes on high speed.

                   Transfer the liquid into a new 15 mL tube. Do not let the stuff on the bottom fall into the new
                   tube. If necessary, use a clean pipette to transfer only the liquid.


                   Slowly add about 4 mL of cold 95% isopropyl alcohol or ethanol to the
                   liquid in the new tube.

                   Mix well by rocking the tube gently back and forth until the DNA becomes
                   visible. It should look like fine white fibers or lint.

              11. Upload the file(s) here (gif,jpg,doc,xls,ppt)

              12. Contact Information

                   First Name: Lori

                   Last Name: LaBorde

                   Email Address: lori.laborde@gmail.com

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              NYU Steinhardt School of Culture, Education, and Human Development: Science Lesson Plan Submission




Page 3 of 3                                                        Report generated by SurveyGizmo on Dec-19 '08

								
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