Amgen Biotech Lab Series

1. Amgen Biotech Lab Series

3. LAB 1: practice micropipetting & loading a gel

4. Micropipet Use • . Twist dial to desired volume • . Add disposable pipet tip • . Press plunger to first stop • . Insert pipet tip into solution to be • transferred • . Slowly release plunger to retrieve liquid • . Move pipet tip into desired tube • . Press plunger past first stop to second stop to transfer liquid, keep the plunger down as you remove it from the tube. • 8. Eject tip Watch “using a micropipettor” Animation in “Animation” File

5. Hold micropipet and epitubes at eye level

6. Loading a gel • Micropipette desired amount • Place tip ABOVE the well in the gel (but under the buffer) • Push through to the 2nd stop • Keep plunger depressed while you pull the micropipette out of the gel box • Write down which well you loaded on the handout taped to the lab bench Do NOT pierce through the bottom of the gel!

7. Micropipet tip should be ABOVE the well NOT IN IT!!!!

8. Micropipetting technique A

9. technique B Close the tips!

10. Technique C Billiard style

12. Micropipet tip punched right through the gel See dye under the wells

13. NICE!

14. After you load the gel: • Throw out dirty tips (keep waste beaker) • Refill tip box with new tips • Return 1, 2, 3 tubes to teacher. • Put 2 new Ependorf tubes in your tube rack for Lab 2a.

15. Gel Electrophoresis separates molecules by: • Size • charge

16. Lab 1 • uses 3 different colored dye molecules that are different sizes and different charges • The 1, 2, & 3 lane will either be 1 type of dye or a mixture of 2 or 3 dyes. • What does the gel below show? • Lane 1: DNA ladder • Lane 5: 397bp vs 187bp Watch “loading a gel” and “agarose run” Animations in “Animation” File