Running head: HONEYBEES DNA BARCODING IN FLORIDA 1 3 HONEYBEES DNA BARCODING

Running head: HONEYBEES DNA BARCODING IN FLORIDA

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HONEYBEES DNA BARCODING IN FLORIDA

Honeybees Dna Barcoding In Florida
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Abstract

The existence of many species of bees in the world calls for DNA extraction to determine the specific types and cycles. For instance, more than 20,000 species of bees exist in the world. Classifying each of the species has been one of the most challenging processes. There is a need for DNA extraction, DNA cleaning, and concentration. These three steps play an essential role in determining the features and impact of each of the species. Many ways have been innovated; however, DNA extraction is one of the best and most scientifically confirmed ways of determining the tests. Despite the existing critiques about the process, it is estimated that it gives the most accurate results of the whole process. Thus, creating a better focus of the entire process through the procedure.

The research experiment will involve the background research, procedure, methods, discussion and conclusion. The background or introduction will give an overview of the whole experiment. The procedure and methods will highlight the major ways the study was conducted. Data was collected, and the DNA extraction data were recorded and analyzed based on the objectives. As per the data collected, the procedure had the DNA nucleotide bases (dNTPs) applied to come up with better conclusions at the end of the research study process. The major objective was to have a DNA extraction that would differentiate the existence of various species of bees. Despite the bee species, it was essential to study the features of the types of bees that exist and their features. Therefore, the study has played a vital role in determining the DNA of the honeybees and other species of insects in Florida.

Introduction

The report has covered the essential elements of DNA extraction, cleaning and concentration in the experiment of a honeybee.  The experiment provides an overview of the whole DNA extraction process considering the aspects of cleaning and concentration. The essential components and elements needed for the process have been stated to help the research reduce the bias that may arise during the study. The PCR process, which plays an essential role in DNA extraction and isolation, has been considered among the significant procedures in the whole experiment. The major equipment and requirements will be listed in the procedures and lab process of the entire experiment. The experiment will also be conducted based on the initial objectives of the research. As such, it is targeting the most accurate results that can be applied in future research and implementation of the reasonable procedure.

The Zymo Research Quick-DNA Tissue/Insect Miniprep Kit will be applied in the DNA extraction process because of its efficiency and accurate results. The kit has been confirmed by many researchers in the genetic engineering field. As a result, the kit fits most of the requirements and objectives of this study. Therefore, it is significant for the researcher to consider the objectives and the final elements of the experiment. The experiment will require 50 grams of been as a specimen. The specimen will be tissue extracted from the bee for the DNA test process. The following steps will be followed in the study;

Preparation of the bee specimen.

Cell Lysis.

Separating the Protein from the cellular debris.

Binding the DNA.

Methods

The study was conducted in experimental research. The experiment involved four significant processes: preparing the specimen, cell lysis, separating the protein from cellular debris, and DNA binding. The use of the been specimen experimented. As such, the following is a detailed explanation of the steps and procedures that were followed in the experiment.

Preparation of the bee specimen.

The specimen needed to be prepared to ensure that it was ready for the DNA extraction process. Also, the preparation process enabled the research to reduce errors because the specimen can be contaminated or dirty. It was essential for the specimen to be rinsed to remove the ethanol that might be excess. Also, the sample must be dried to enhance visibility and ensure that it could yield the required results. The bee is required to be dissected into two equal half where only one segment could be used in the experiment. Finally, the bee was taken to a lysis tube to maintain the tissue part of the specimen. The process must be done because it acts as a way of checking the essential features of the specimen to be used (Hajibabaei et al., 2017). Therefore, the specimen has to be well prepared.

Cell Lysis

The specimen will be mainly separated by grinding and mixed in a saline solution. At this, the major aim is to protect the negative phosphate ions that are essential in the DNA extraction process. As such, the negative sodium ions will play a significant role in protecting the phosphate ions for the experiment’s success. Also, more detergents are needed to eliminate the fat layers from the tissues used as specimens. Therefore, the procedure was mainly conducted to ensure that the molecules could move freely to make it easier for the DNA extraction to be easily conducted.

Separating DNA from the Protein Debris

The procedure was essential as it was meant to get rid of the cellular debris to increase the accuracy of the experiment. Besides, it was meant to reduce the excess unwanted components in the DNA extraction process. The first step was to apply a protein enzyme to reduce the excess fats in the cells. Finally, the sample was filtered, and only the wanted components could be applied. The filtered sample is pure and has no impurities that will tamper with the whole process.

Binding the DNA

Binding the DNA was one of the most important processes in the whole experiment. More significantly, the stage helped isolate the DNA into the silicon column (Brown et al., 2017). This is the last process of DNA extraction; the next step will be the kit exaction to identify the essential features and components of the bee specimen. Therefore, it was easy to conduct the purification process.

Results

The study results were displayed in the DNA universal primer scales; the scales could present the nature of the data collected. The scaling level could determine the extraction conducted and display the results as per the expected analysis. The collected results could be compared with the normal values on the scales. The results were posted in four significant strands of DNA that had different scaling. The different scaling created a better way of assessing the genetic composition of the elements. As such, each strand had a different composition based on the structure. Therefore, the following interpretations are made on the DNA extraction and cleaning process.

 

Figure 1: Universal Primer1

 

Figure 2: Universal Primer 2

 

Figure 3: Universal Primer 3

 

Figure 4: Universal Primer4

Discussion

The research experiment played a significant role in assessing the DNA of the honeybee. Especially in the process of extracting the DNA samples from the kit. Despite the considerable challenge of time and some of the processes having challenges during the process, many observations were noted. The denaturing process was one of the critical sections and the center of discussion based on its impacts on the data set. As such, based on the data collected, there were four strands of the DNA extraction process of floral honey (Hawkins et al., 2017). The process involved heating in extreme high temperatures to form the next building blocks round of extraction. The section of the results displayed by the four different primers showed a different genetic composition because of the different scales.  Therefore, this implies the honey has four different strands of DNA scales that could yield multiple genetic compositions, as shown in the figures posted under the results.

Annealing and extending also played a significant in the experiment. For example, they could easily ensure that the temperature was lowered to carry out the other experiment. More significantly, the DNA primers will easily attach themselves to the DNA template. The temperature will also be raised to ensure that the process is repeated until the last finding is collected. The results collected will affirm the existence of DNA purity in the sample under extraction. The essential component was the kit and the other elements that could create accurate results in the experimental part (Packer et al ., 2017). Therefore, the honeybee DNA was represented by the four DNA strands extracted in the process. Thus, the four strands could be studied, and better conclusions made about the whole and its implications at the end. 

References

Brown, M. J., & Paxton, R. J. (2017). The conservation of bees: a global perspective. Apidologie, 40(3), 410-416

Hajibabaei, M., Singer, G. A., Hebert, P. D., & Hickey, D. A. (2017). DNA barcoding: how it complements taxonomy, molecular phylogenetics and population genetics. TRENDS in Genetics, 23(4), 167-172,

Hawkins, J., de Vere, N., Griffith, A., Ford, C. R., Allainguillaume, J., Hegarty, M. J., … & Adams-Groom, B. (2017). Using DNA metabarcoding to identify the floral composition of honey: A new tool for investigating honey bee foraging preferences. PLoS One, 10(8), e0134735.

Packer, L., Gibbs, J., Sheffield, C., & Hanner, R. (2017). DNA barcoding and the mediocrity of morphology. Molecular Ecology Resources, 9, 42-50.