Introduction:

In this lab, we will be isolating DNA from mouth swabs using a provided kit.
Materials/Methods:
Materials Kit Materials
Buccal swabs Lysis buffer (L11)
Magnarack Proteinase K
Sterile 1.5mL micro tubes Magnetic beads
Vortex mixer Purification buffer (N6)
Sterile pipette tips (20ul, 200ul, and 1ml) Wash buffer (W12)
Water bath Elution buffer (E5)

Before starting
1. Set water bath 37c
2. prepare lysis mix
a. mix 1ml of L11 and 10ul proteinase K
3. vortex tube containing magnetic beads to resuspend
4. prepare purification mix
a. mix 40ul magnetic beads + 100ul N6
Preparing lysate
1. transfer buccal sample to sterile micro tube
2. add 1mL lysis mix
3. incubate in bath for 20min
Binding DNA
1. transfer the digested supernatant into new tube
2. pipet up and down purification mix with magnetic beads
3. add 140ul of purification mix to sample and mix gently
4. incubate at room temp for 1min
5. place sample onto magnarack for 1min
6. remove tube from magnarack
7. remove the supernatant and discard
Washing DNA
1. add 1 ml of W12 to sample
2. leave sample on magnarack for 1min
3. remove tube from rack and discard supernatant
4. repeat 1-3
Eluting DNA
1. remove tube with pelleted beads from rack
2. add 150ul of E5 to tube and pipet up and down to resuspend beads
3. incubate for 1min at room temp
4. place on rack for 1min
5. remove tube and remove supernatant containing DNA to sterile micro tube
6. DNA can be stored at -20C.