INTRODUCTION

It has long been established that the deoxyribonucleic acid (DNA) is responsible for the traits and
characteristics expressed by an organism. However, small changes in the DNA sequence can lead to dire
effects in the organism like malfunctioning systems and diseases.

In a eukaryotic cell, the DNA is organized into chromosomes, which is a package of DNA with protein
complexes. These chromosomes occur in homologous pairs in the cell, meaning for each DNA sequence,
the cell has two copies of it, except for its sex chromosomes labeled X from the female and Y from the
male. By observing chromosome pairs, aberrations or abnormalities in the chromosomes can be
pinpointed and traced back to the mutation in the DNA such as deletions, duplications, inversions and
translocations (Alberts et al, 2008).

Errors in the DNA are found through genetic testing. Different tests that look for anomalies in the DNA are
available nowadays due to technology but the most basic of them is karyotyping. A karyotype is a
photomicrograph of a cell’s chromosomes arranged in pairs based on the size, centromere position and
banding pattern (due to staining) of the chromosomes (Starr and Taggart, 2006). The pairs in the
karyotype are presented from the largest to the smallest and are numbered from one to so on with the
exception of the X and Y chromosomes (Brooker, 2005).

A karyotype is usually obtained by first stimulating cell division. During cell division, the chromosomes
shortens and become more tightly coiled making their shapes more distinct and hence more visible under
the light microscope especially during the metaphase stage of division. After which, the cells are arrested
at metaphase through the addition of colchicine which prevents the formation of spindle fibers and so
halts mitosis. Then the mixture is centrifuged. The resulting pellet is immersed in a hypotonic solution that
causes the cell to swell and so allows more space for the chromosomes to spread. Afterwards, a fixing
agent is applied on the cell to freeze the chromosomes from moving. A stain is subsequently employed to
allow visualization of the chromosomes and their banding pattern. A photograph of the chromosomes is
then taken, cut, paired and rearranged to give a karyotype (Starr and Taggart, 2006).

Through the karyotype, the banding pattern of the chromosomes is displayed. The banding pattern
enables the researcher to recognize specific parts of the chromosome and to identify parts of the where
deletions and translocations have occurred (Lodish et al, 2000). The chromosomes can also be
differentiated from each other and changes in its structure are detected. Since they can be visualized,
similarities between chromosome appearances among species are observed and thus help in determining
the evolutionary relationship of species (Brooker, 2005). The number, shapes and sizes of the condensed
chromosome vary for each species (Lodish et al, 2000) hence, closely related species can be
distinguished from each other (Brooker, 2005).

In the laboratory, the actual experiment made use of onion root tips as the karyotype source. The method
employed is different and less complex from the one stated above since this is a plant source. First the
onion roots were grown in water and its tips were subsequently cut off and fixed in 3:1 ethanol: glacial
acetic acid. The sample was then stained with acetocarmine dye in acetic acid and was subjected under
the squash method (“Acetocarmine Staining,” 2007). The squash method makes use of the thumb to
apply vertical pressure on the sample to make it thinner and flatter for viewing under the compound light
microscope. In doing so, the specimen appears to be more transparent and so light can easily penetrate
through it allowing for a better visualization of the cells and the chromosomes. When this was done, the
cells were photographed and documented.

By using actively dividing cells in the onion root tip, this experiment aims to obtain a karyotype from the
sample and to determine the purpose of each step used in the procedure.
References:

“Acetocarmine Staining.” Wheat Genetic and Genomic Resources Center. 26 April 2007. Kansas State
University. 25 August 2008 <http://www.k-state.edu/wgrc/Protocols/Cytogenetics/acetocarmine.html>.

Alberts, Bruce, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts and Peter Walter. Molecular
Biology of the Cell. 5th ed. New York: Garland Science, Taylor and Francis Group, LLC, 2008.

Brooker, Robert. Genetics: Analysis and Principles. New York: McGraw-Hill Publishing, 2005.

Lodish, Harvey, Arnold Berk, S. Lawrence Zipursky, Paul Matsudaira, David Baltimore and James Darnell.
Molecular Cell Biology. 4th ed. New York: WH Freeman and Company, 2000.

Starr, Cecie and Ralph Taggart. Cell Biology and Genetics. California: Thomson Brooks/Cole, 2006.