Safari Park Condor Lab Procedures

Safari Park Condor Lab Procedures

Pre-lab questions:

1.  Draw a labeled diagram of a micropipette and explain how it is used.

2.  Why is PCR considered to be DNA replication in a test tube?

3.  How specifically is PCR being used in this lab?

4.  Explain how to identify the gender of a condor from its karyotype.

5.  What is the relationship between the condor karyotype and the gel electrophoresis results?

6.  Evaluate the importance of DNA biotechnology in the effort to conserve the California condor.

Day 1: PCR

1.  Label the top of your tube with a permanent marker. Make a note of how you labeled your tube in your notebook for future reference.

2.  Add 18 ul of primer mix and 2 ul of condor DNA to the PCR tubes. (Note: the PCR tubes have a blue solution at the bottom that contains free nucleotides, the building blocks of DNA.)

3.  Mix the contents of the tube by flicking gently with your finger (see teacher demo).

4.  Centrifuge the contents of the tube.

5.  Place the tube into the thermal cycler (PCR machine).

6.  Your teacher will turn on the thermal cycler to replicate your condor DNA in vitro.

Day 2: Gel Electrophoresis

1.  PREPARE THE GEL RIG (Performed by TEACHER)

a.  Assemble the gel rig by connecting the power strip (with AC/DC switching adaptor) to the E-Gel iBase. Remove SYBR Safe gel from packaging, and slide into position on the E-Gel iBase. Press the “Mode” button and use the up and down arrows until “0 PRE-RUN 2:00” is selected. Press the “Go” button. When the pre-run is finished, press the “Go” button again. Gently remove comb from the gel.

2.  LOAD PCR PRODUCTS (performed by STUDENTS)

a.  Load all 20 uL of your PCR products into individual wells in the gel.

Teacher note: If there are any empty wells after all students have loaded, load 20 uL of sterile water into these.

3.  RUN THE GEL (Performed by TEACHER)

Press the “Mode” button and use the up and down arrows to scroll until “1 E-Gel 0.8-2% 35:00” is selected. Press the “Go” button.

4.  VIEW THE GEL (Performed by STUDENTS)

a.  To stop the run, press the “Go” button.

b.  Place the UV shield on top of the E-Gel iBase and nest entire unit on the E-Gel Safe Imager, and connect the Safe Imager to the iBase with attached cord.

c.  Press the red button at the front of the E-Gel Safe Imager to turn on the UV light.

d.  If viewing is difficult, take the entire gel rig into a dark room or closet to view.

5.  INTERPRET RESULTS

a.  There may be other banding present, but you should be able to identify the Z and/or W bands as the bands that moved down the farthest in each well (see example gel image below).

b.  Use the banding pattern to determine if the condor is male or female (one band: male, two bands: female).

Reflection questions:

1.  What did you learn?

2.  What was your favorite part of the lab?

3.  Draw a picture of your lane on the electrophoresis gel.

4.  What was the sex of your condor? How do you know? Explain.