Biology Wed May 18, 2005

Homework:

Warm Up:

1. Write out a set of crystal clear numbered instructions for HOW TO USE A MICROPIPET.

2. Each restriction enzyme recognizes and cuts at (one/many different) restriction site(s).

3. Small circular rings of DNA in a bacterial cell are called (sticky ends/ plasmids).

4. Plasmids (can/ cannot) move in and out of a bacterial cell.

5. Why in LAB B, was cabbage juice added to the gel box?

6. A sticky end is (double / single ) stranded DNA.

7. The DNA molecules has a (positive / negative ) charge.

8. What does buffer do? Why will we use it?

9. Write a palindrome.

10. Use the following terms to describe how ganetic engineers create a bacterium that produces a human protein product such as insulin.

ligase restriction enzyme

restriction site plasmid

protein product made sticky ends

recombinant plasmid DNA containing the human gene of interest.

Hold micropipet

and epitubes at

eye level

Micropipet UseMicropipet Use1 Twist dial to desired volumeTwist dial to desired volume2 Add disposable pipet tipAdd disposable pipet tip3 Press plunger to first stop Press plunger to first stop 4 Insert pipet tip into solution to be Insert pipet tip into solution to be transferredtransferred5 Slowly release plunger to retrieve liquidSlowly release plunger to retrieve liquid6 Move pipet tip into desired tubeMove pipet tip into desired tube7 Press plunger past first stop to second stopPress plunger past first stop to second stop to transfer liquid, keep the plunger downto transfer liquid, keep the plunger down as you remove it from the tube.as you remove it from the tube.8. Eject tip8. Eject tip

Micropipetting

technique A

technique B

Close the tips!

Technique

C

Billiard style

Micropipet tip should

be ABOVE the well NOT IN

IT!!!!

Micropipet tip punched

right through the

gel

See dye under the

wells

NICE!