Alu PCR Lab
Objective
1. To successfully isolate DNA from, cheek cells.
2. Prepare a PCR reaction for amplification of an Alu instert
Materials
.9% saline solution
micro pipettes, tips
waste container
micro centrifuge
micro centrifuge tubes
PCR tubes
agarose
1x TAE
gel chambers and molds
lead dye
chelex
racks primer mix
master mix
H20
positive control DNA
Procedure
- Swirl 10 mL of saline solution in your mouth for 30 seconds.
- Expel saline into a cup to mix cells
- Label a tube and transfer 1000 micro liters of the saline solution into the microfuge tube
- Spin the saline cell solution in a microcentrifuge to pellet the cells
- Pour off the supernatent into the your cut with about 100 micro liters left in the tube
- Rack or flick the tube to resuspend the cells
- Obtain a tube of 50 micro liters chelex and transfer the cells in the tube before to the chelex tube
- Put the chelex tube into the heat block for 10 minutes
- After heating release the pressure and resuspend the tube
- After spin the chelex for 1 minute
- Obtain another microfuge tube and withdraw 50 mL of the supernatent from the chelex tube
- Put the tube in a rack for the teacher to refrigerate them
- Obtain a tiny PCR tube and pipet 20 micro liters of the master and primer mix along with 10 micro liters of your DNA
- Spin the tube and add 5 micro liters of loading dye into your PCR tube
- Dilute 40x of Agarose and top of the graduated cylinder with water
- heat until dissolved and wait until cool to poor the mixture into the mold
- Load 15 to 20 mL into your gel
- Run the gel
Objective
1. To successfully isolate DNA from, cheek cells.
2. Prepare a PCR reaction for amplification of an Alu instert
Materials
.9% saline solution
micro pipettes, tips
waste container
micro centrifuge
micro centrifuge tubes
PCR tubes
agarose
1x TAE
gel chambers and molds
lead dye
chelex
racks primer mix
master mix
H20
positive control DNA
Procedure
- Swirl 10 mL of saline solution in your mouth for 30 seconds.
- Expel saline into a cup to mix cells
- Label a tube and transfer 1000 micro liters of the saline solution into the microfuge tube
- Spin the saline cell solution in a microcentrifuge to pellet the cells
- Pour off the supernatent into the your cut with about 100 micro liters left in the tube
- Rack or flick the tube to resuspend the cells
- Obtain a tube of 50 micro liters chelex and transfer the cells in the tube before to the chelex tube
- Put the chelex tube into the heat block for 10 minutes
- After heating release the pressure and resuspend the tube
- After spin the chelex for 1 minute
- Obtain another microfuge tube and withdraw 50 mL of the supernatent from the chelex tube
- Put the tube in a rack for the teacher to refrigerate them
- Obtain a tiny PCR tube and pipet 20 micro liters of the master and primer mix along with 10 micro liters of your DNA
- Spin the tube and add 5 micro liters of loading dye into your PCR tube
- Dilute 40x of Agarose and top of the graduated cylinder with water
- heat until dissolved and wait until cool to poor the mixture into the mold
- Load 15 to 20 mL into your gel
- Run the gel