Biology Lab: Simple DNA Extraction

Introduction: All living cells contain DNA. DNA is composed of long chains of genes. Genes contain the
protein building instructions for the cell. It was first determined in the 1940 s that DNA was the carrier of the genetic information! but it was not until 19"# that $atson and %ric& determined the structure of the DNA molecule. 'oda(! much is &nown about how DNA codes and replicates information! and how that information is e)pressed in the phenot(pe of an organism. In this lab! we will attempt to remove the DNA from the cells of wheat germ. $heat germ is composed of the embr(onic cells of wheat seeds. $arm water will be used to soften the plant cell walls and ordinar( dishwashing detergent is used to brea& up the membranes! allowing the DNA to precipitate in the solution.

Problem: $hat does DNA loo& li&e* %an DNA be e)tracted from the nuclei of eu&ar(otic cells* Materials Needed:
$heat Germ "0 m, bea&er $arm water bath +eral pipet Ice cold eth(l alcohol +alance $ood splint Graduated c(linder Dawn dishwashing detergent -icroscope and microscope slides

Safety Precautions:
1. /. ,aborator( aprons and safet( goggles are re.uired for this lab. Alwa(s use caution when using laborator( chemicals. Never touch! taste or smell chemicals unless instructed to do so.

Procedure:
1. /. #. 0se the balance to weigh out appro)imatel( one gram of wheat germ. 1lace the wheat germ in a small "0 m, bea&er. ,ocate the water bath. 'his water is being &ept at a temperature between "0 2 30 %. $ith a small bea&er! dip a small amount of the water out of the water bath. -easure /0 m, of the warm water with a graduated c(linder. 1our the warm water into (our test tube or bea&er containing the wheat germ. 4. 0sing a wooden splint or glass stirring rod! mi) constantl(! but gentl(! for three minutes. ". 0se a graduated +eral pipet to obtain 1 m, of Dawn dishwashing detergent. 4elease the detergent into the bea&er with the wheat germ. 3. 5tir gentl( for " minutes! being careful not to create any foam. 6If (ou do create foam! use a dropper or pipet to remove the foam.7 8. 0se a graduated c(linder to measure 1" m, of ice9cold eth(l alcohol. :. ;old the bea&er of wheat germ at a 4"9degree angle! and very slowly pour the alcohol down the side of the bea&er. If poured correctl(! the alcohol will form a la(er on top of the wheat germ and detergent mi)ture. Do not mi) the la(ers together. 'he DNA will precipitate at the boundar( between the alcohol and the water. If the water la(er and the alcohol la(er mi) together! the DNA will not precipitate. 9. Do not move the bea&er for at least 1" minutes. $hite! string( filaments of DNA will begin to appear at the water < alcohol boundar(. If (ou let the solution sit undisturbed for 1" minutes! the DNA will begin to float to the top. 10. 0se the wood splint or glass stirring rod to begin to spool the DNA. =er( gentl( swirl the wood splint through the DNA. >ou should be able to wind the DNA around the wood splint. 11. 'o get more DNA to precipitate! dip the wood splint ver( gentl( down into the water la(er! lifting the water up into the alcohol la(er. 1/. 1lace a small amount of (our DNA on a clean microscope slide. Add a drop of meth(lene blue and a coverslip. ?bserve under low and high power.

bser!ation "uestions:
1. $hat purpose is served b( soa&ing the wheat germ in the warm water*

/.

$hat purpose is served b( the Dawn dishwashing detergent*

#.

Describe what the DNA loo&s li&e with (our na&ed e(e.

4.

Describe what the DNA loo&s li&e under the microscope.

".

$here did the DNA first appear*

3.

$hat might have happened if (ou poured the alcohol too fast and it did not form a la(er on top of the water*

8.

'here is about 3 feet of DNA contained in each of (our cells. ;ow is it possible for so much DNA to fit inside the small space of a cell*

:.

$h( is DNA referred to as a double heli)*

9.

$hat are the building bloc&s that compose DNA* $hat are the three components to these building bloc&s*

10. Draw a strand of DNA that has the following base se.uence on the left hand side@ A''%G%'%G. 0se the base paring rules to construct the right hand side of the molecule. ,abel the location of the sugars! the

phosphates! and the nitrogen bases. Indicate where (ou would find a h(drogen bond.

11. DNA is able to ma&e an e)act cop( of itself. $hat is this process called* $h( is it so important that DNA be able to ma&e an exact copy of itself*

1/. $hat role does each of the following enA(mes pla( in the replication of DNA* a7 DNA pol(merase

b7 ;elicase

#eac$er Preparation:

1.

'his is a B.uic& and dirt(C DNA e)traction. 'here are man( other lab procedures for this that can be used to e)tract a purer sample of DNA. I li&e this procedure for m( standard +iolog( I students! because it is simple and straightforward. It can be done .uic&l(! with minimal lab materials! and students can easil( e)tract the DNA. 'he temperature of the water is ver( important. It must be &ept between "0 2 30 %. 'ools that can be used to spool the DNA@ 1aper clips! wood splints! bamboo s&ewers! glass stirring rods. It is essential that the alcohol be ice cold. I &eep it in the freeAer prior to the lab. During the lab! I &eep it in an ice buc&et. ;ave students tr( a variet( of other cell materials and compare them. I have also used strawberries! onions and liver. I have tried different brands of dishwashing detergent! and BDawnC alwa(s wor&s the best. Dth(l alcohol seems to give me the best results! but I have used isoprop(l alcohol and this also gives good results.

/. #. 4. ". 3. 8.

Ans%ers to "uestions:
1. /. #. 4. ". 3. 8. :. 9. 10. $arm water is used to soften the plant cell walls. Dawn dishwashing detergent is used to brea& up the membranes! allowing the DNA to precipitate in the solution. 5tudent answers will var(. 5tudent answers will var(. 'he DNA will first appear at the boundar( between the warm water<detergent solution and the cold alcohol. If the alcohol is added too fast! the water and alcohol will mi) together. If the water la(er and the alcohol la(er mi) together! the DNA will not precipitate. 'he DNA is tightl( wound around proteins called histones. ;istones are then supercoiled into nucleosomes. DNA is referred to as a double heli) because it is two sided and twists around in a spiraling fashion. DNA is composed of monomers called nucleotides. A nucleotide is composed of three parts@ a sugar molecule! a phosphate molecule! and one of four nitrogen bases.

11. $hen DNA ma&es an e)act cop( of itself! it is called replication. 4eplication occurs prior to mitosis and

meiosis. An e)act cop( of all chromosomes must go into new daughter cells. 1/. DNA pol(merase catal(Aes the elongation of new DNA b( the addition of new nucleotides at the replication for&. ;elicase is the enA(me that untwists the double heli) of DNA at the replication for&s! separating the two strands and ma&ing them available as template strands.