DNA Isolation Lab. COLLECT AND LYSE CELLS Fill 15 mL test tube with 3 mL of water Label with your...

DNA Isolation Lab

COLLECT AND LYSE CELLS

Fill 15 mL test tube with 3 mL of waterLabel with your initials.

GENTLY chew on the inside of your cheek to loosen cells.

Images from BIO-RAD lab manual

COLLECT AND LYSE CELLSUse the 3 mL of water in tube to rinse your mouth and SPIT liquid into 15 mL tube or cup.DON’T SWALLOW LIQUID!

http://www.scienceprogress.org/2008/10/spitomics/

COLLECT AND LYSE CELLS

Using a new clean pipette,ADD 2 mL of LYSIS BUFFERto your 15 mL tube.

Place cap on tube. GENTLY INVERT 5 TIMES to lyse cells. DON’T SHAKE THE TUBE!


REMOVE PROTEINS

Add 5 DROPS of PROTEASE + SALT solution.

Place cap on tube. GENTLY INVERT 5 TIMES to mix DON’T SHAKE THE TUBE!


REMOVE PROTEINS

Place tube in 50° C WATER BATHfor 10 minutes to allow protease to work.


http://www.directindustry.com/prod/stuart-equipment/laboratory-test-tube-block-heaters-63431-412378.html

http://img.directindustry.com/images_di/photo-g/laboratory-test-tube-block-heater-412378.jpg

MAKING THE DNA VISIBLE

Fill a disposable pipette with ICE COLD ALCOHOL.

Hold tube at a 45 degree angle. Add alcohol by SLOWLY dispensing it down the inside wall of the tube so it forms a layer on top. It will take repeated additions to add 10 mL.


Screw cap on tube and place upright in test tube rack.

Leave it undisturbed at room temperature for 5 minutes.

After 5 minutes look at tube (especially in area where alcohol and cell extract layers meet)

http://www.arssales.com/equine/assets/images/test_tube_rack.jpg

http://learn.genetics.utah.edu/content/labs/extraction/howto/


After 5 minutes look at tube (especially in area where alcohol and cell extract layers meet)

TIGHTLY CAP TUBE and SLOWLY INVERT 5 times.

.

The stringy white material is DNA.The salt you added helps it stick togetherDNA stays dissolved in water, but when salty DNA comes in contact with alcoholIt “undissolves”.


Use a disposable pipette totransfer your DNA into a vial alongwith 750 uL – 1 mL of alcohol.

Dry off the opening of the tube andseal with “Crazy glue”

http://paint-and-supplies.hardwarestore.com/50-272-super-glue.aspx

DNA Isolation Lab. COLLECT AND LYSE CELLS Fill 15 mL test tube with 3 mL of water Label with your...

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