1. Biotechnology
Genetic Testing

2. Micropipetes
Liquid measurements in the metric system are made in units based on the liter where a liter is about one quart.
To make these precise measurements, molecular biologists use a precision tool known as a micropipet. This tool is as basic to their lab work as a hammer is to a carpenter.
Micropipets come in many models and sizes. You will be using micropipets similar to those found in the Fred Hutchinson Cancer Research Center research labs

3. General Guidelines
Set the volume only within the range of your micropipet.
Have the proper size disposable tip in place on your micropipet before immersion into any solution.
Always keep the micropipet in a vertical position when there is liquid in the tip. In a horizontal position, fluid can leak back into the piston.
Use your thumb to control the speed at which the plunger rises after taking up or ejecting liquid. Letting the plunger snap back damages the piston and the volume dispensed.

4. How to use Micropipete Set amount Put on tip Press down to first stop
Put into solution
Release
Go to location
Go down to second stop
Tip??

5. You try on paraffin– everyone at team needs to do one!
1ul
5ul
10ul
20ul
100ul
500ul
Damp table will make less curl of paraffin

6. General Guidelines
Set the volume only within the range of your micropipet.
Have the proper size disposable tip in place on your micropipet before immersion into any solution.
Always keep the micropipet in a vertical position when there is liquid in the tip. In a horizontal position, fluid can leak back into the piston.
Use your thumb to control the speed at which the plunger rises after taking up or ejecting liquid. Letting the plunger snap back damages the piston and the volume dispensed.

7. How to use Micropipete Set amount Put on tip Press down to first stop
Put into solution
Release
Go to location
Go down to second stop
Tip??

8. Hints: Gel with wells + buffer on top
Set amount  put on tip
Load solution (see yesterday’s notes)
Tilt pipet a little  use two hands – Ms.P grabs wrist.
Break through buffer, but do not stab into Gel – sample will sink into well (ok today, much better on Thursday)
Stop at 1st stop  ok if some of sample left inside pipet – do not want to get air bubbles

9. Practice Gels!
Each person at team must pipet in 15ul of solution (any color is fine) into a well
Repeat!
P10: ,
P20: 15

10. Gel Boxes: how work? 1. Charges: Uses Current to “pull” molecules
Black = negative = so positive will be attracted to it
Red = positive = so negative will be attracted to it
2. Size:
Larger objects move slower through the gel
Smaller objects move faster through the gel

11. Data Table Observations - Red pH - Red Observations - Black pH - Black
Water + NaCl + Phenol Red
Buffer + Phenol Red

12. Gel Boxes and Power Sources
See sheet for voltage amount
TURN OFF when disconnecting cords!
Gel Boxes and Power Sources
Add 125ml distilled water + 250ul NaCl + 1ml Phenol red – mix well – turn on to ~100v – record observations + pH at both ends (red and black)
Turn off  take out tray  dump down sink  dry  put back in box
Add 125 TAE buffer + 1ml phenol red – mix well – turn on to ~100v – record observations + pH at both ends (red and black)
Turn off  take out tray  dump down sink  dry 

13. Data Table Observations - Red pH - Red Observations - Black pH - Black
Water + NaCl + Phenol Red
Yellow/clear color – bubbles by metal bar
5-6
Bright pink, fuchsia – bubbles by metal bar
11-9
Buffer + Phenol Red
5-7

14. Molecule movement – Dyes to mimic DNA
Pipet in 20ul of each dye into your gel (wells are in the center) – each team member does one – bring notes
Person 1 and 2, Person 3 and 4, Person 5 and 6
Person 6: Set box to 100v  run for 10 min (make sure to wait for partner box before using power x
Open lid  take a picture of your gel
First person that pipetted will take picture can clean up

15. Data Table Well number from top of box
Number of dye (on top of microtude)
Color of dye before
Dye migrated to black or red end
Color of dye after
Order of distance from center (1st closest to center, 8 is furthest) 1 2 3 4 5 6 7 8

16. Underneath/Next to your table...
Draw DNA (backbone and rungs) with the following strand (you need to come up with the complementary strand): A T C G

17. Post-Lab Questions

18. Post-Lab Questions
Which dye molecules (give #) had a positive charge? Explain.
Which dye molecules (give #) had a negative charge? Explain.
Which dye molecule was the largest? Explain.
Which dye molecule was the smallest? Explain
How could scientists and doctors use Gel electrolysis to analyze someone’s DNA?