Report on Chicken Gender Determination via DNA Extraction and Analysis

This report details an experiment on determining the gender of chickens through DNA extraction from feather samples. The introduction highlights the challenges of DNA isolation, emphasizing the importance of choosing the appropriate method based on the organism and tissue type. It references various methods for DNA extraction from animals, including blood, feathers, and tissue, and the use of methods like spectrophotometry and gel electrophoresis for analysis. The experiment focuses on using feather samples and the DNeasy Blood and Tissue kit to extract DNA, followed by analysis using spectrophotometry and gel electrophoresis. The results section presents data on the A260/280 ratio and concentration of the extracted DNA, along with observations from gel electrophoresis, showing the success of DNA extraction from feathers. The report discusses the methods used, including the collection of feathers, DNA purification with the Qiagen DNeasy Blood & Tissue kit, and subsequent analysis. It also acknowledges the limitations of using feather samples due to the lower DNA content compared to blood or tissue, and the importance of proper DNA storage. The conclusion reiterates the necessity of DNA extraction for molecular biology applications, the importance of DNA quality and purity, and the efficiency of the methods employed. The report provides a comprehensive overview of the procedures, results, and analysis involved in the study, offering insights into gender determination in chickens through DNA extraction from feathers, and provides a valuable resource for students on Desklib.

Genetics and Genomics
INTRODUCTION
Isolation of DNA is however a quite challenging it depends upon the type of organism and the type of tissue used. As variability in DNA impacts the results of an outcome therefore it becomes necessary to choose an good or best
DNA isolation technology. There are various methods which is used in order to extract DNA from animals. However sample of DNA is collected by hair, blood, feathers, faeces, vomit, saliva, bone, teeth and tissue. These are some of the
ways which are used in order to collect DNA of animals.
As per the views of M Ghaheri & et.al., (2017), every organism consist of DNA. (Deoxyribonucleic acid) in their cells. It is a complex molecule which consist of all information which is necessary in order to maintain organism. However
extraction of blood for sample is one of the most common method which is used. The DNA which has been extracted is then analysed as per the demands, quantity and quality. And the extracted DNA is surveyed by use of nano drop
spectrophotometry method and gel electrophoresis. There is no significant difference between two of the methods of DNA purity. Other than this phenol/chloroform are the most toxic method and takes more time but is higher than other
methods (Vilstrup, J. T. & et.al., 2018).
In this experiment various methods will be used in order to determine the gender of chicken as it is very difficult to know the gender of chicken the reason behind this can be, in sexually demographic species different genders can
be easily classified while it might not be easy for monomorphic species. Therefore extraction of DNA might be useful in gender determination hence this report will use all the three tissue types for determining sex. be extracted from
feathers.
METHODS
DNA extraction from feather
In order to find the gender of chicken Qiagen Dneasy Blood & Tissue kit has been used in order to purify DNA from feathers. The feathers of chicks are collected and than it analysis has been done. The DNA of chickens has been extracted by
using the above mentioned purification kit. And the DNA which was found has been analysed by using spectrophotometer. The samples which has been collected is then electrophoresed in agarose gel in order to determine intensity of band for
extraction of DNA (Pandey & et.al., 2016).
Then all the results which has been found is compared with class results. In this method as per Vilstrup, J. T. & et.al., (2018), a few feathers are plucked from top of head or breast and it has been placed in ethanol. And then it is stored for few
months at room temperature.
PCR Method
Then PCR method has been used in order to make the copies of DNA
segment. All the ingredients i.e., taq polymerase and PCR primers has been kept in
container and repeated heating to separate DNA and then it is cooled so primer
can bind and again temperature is raised to extend primer and synthesise
new strand (Quick & et.al., 2017).
Gel electrophoresis Method
Then TAE buffer has been prepared by transferring 100ml of buffer abd then 2 gm of agarose is added. After keeping this solution in oven ehtidium bromide has been added and the solution so formed has been kept on comb. Then buffer
solution has been placed in electrophoretic chamber and after connecting electrode switch is on. Then gel has been placed in UV transilluminator.
RESULTS: However extraction of DNA from feather has been proved successful. Figure 1 represents about the gel electrophoresis
which has been used in order to separate and identifying DNA of chicken by using feather to collect sample. It has been done by using
DNeDNA extraction and purification. 2019.asy Blood and Tissue kit. In analysis it has been found that feather sample which has been
extracted is displayed poorly,
After electrophoresis sample has been diluted in concentration. By comparing the bands in sample their approximate size
can be determined. In this process if band is too faint then it is left undiluted. Figure 2 represents gel electrophoresis by extraction of
DNA by feather.
CONCLUSION: DNA extraction is required for molecular biology application. Many kits are available for analysing DNA. The
first and foremost step is to collect a sufficient amount of DNA and then removing contamination from it and lastly ensuring
quality and purity of DNA. The purity of DNA depend upon the type of extraction method used. The DNA is extracted from
animal tissues and cells (Kaufmann & Pitt, 2018). However in study Dneasy blood and Tissue kit has been used. It is used to
isolate DNA from various sources such as blood, feather, tissue, insects, hair etc., however in study feather has been used. The size
of purified DNA is about 50kb in size. The procedure of DNA extraction using this method takes about 20 minutes (A single
plucked feather as a source of DNA, 2019). The method which has been used which is gel electrophoresis and PCR method is
however a proven method for separation of DNA fragments. The DNA has been extracted from blood clot of feather. This data
sampling method is however not as effective as that of other sampling methods. The reason behind this is that feather consist of
typically low DNA in comparison to blood and tissue. Even if the sample provide some result it lacks quality in comparison to
blood sample.
As this experiment has been conducted for a number of week. Therefore it becomes extremely important that DNA has
to be stored in a 4 degree C for weeks, while 20 degree C for months and 80 degree C for years as per the general rule. As keeping
DNA for long may result in damage therefore proper preservation has be done. The DNA has been kept at room temperature and
importance of DNA has also been stated (DNA extraction and purification, 2019).
REFERENCES
Books and Journals
Vilstrup, J. T. & et.al., (2018). A simplified field protocol for genetic sampling of birds using buccal swabs. The Wilson Journal of Ornithology. 130(1). 326-334.
Kaufmann, M. E., & Pitt, T. L. (2018). Pulsed-field gel electrophoresis of bacterial DNA. In Methods in practical laboratory bacteriology. (pp. 83-92). CRC Press.
Quick, J. & et.al., (2017). Multiplex PCR method for MinION and Illumina sequencing of Zika and other virus genomes directly from clinical samples. nature
protocols. 12(6). 1261.
Pandey, U. & et.al., (2016). DNA from dust: comparative genomics of large DNA viruses in field surveillance samples. Msphere. 1(5). e00132-16.
Online
A single plucked feather as a source of DNA. 2019. [Online]. Available through : <https://sora.unm.edu/sites/default/files/journals/auk/v108n04/p0959-p0960.pdf>.
DNA extraction and purification. 2019. [Online]. Available through <https://www.labome.com/method/DNA-Extraction-and-Purification.html>
Initials Sample Code A260/280 Concentration (ng/uL)
DY, S B18 1.723 45.19
T,M B35 2.125 18.38
AJ, RA, ZM B40 1.756 29.7
NS,ZB B49 1.806 40.36
AT, AA B49b 1.89 39.52
AW,JM B51 1.748 25.93
LB,SK B51B 1.799 24.04
MS,LZ B61 1.915 42.06
AV KC F28 2.076 68.18
RA,JD F35 1.322 9.94
SP,R F37 1.557 4.3
AA,ZA F39 1.079 5.52
AP, LW F41 1.439 10.44