Introduction
The dopamine transporter gene in human has had a strong association with certain neuropsychiatric disorders and behavioral conditions (1). This is because neurotransmission of dopamine underlies brain functions such as locomotion, cognition, behavior, and motivation (1). Interference with the production, regulation and signaling of dopamine and its neurotransmissions can cause neuropsychiatric diseases such as attention deficit/hyperactivity disorder (ADHD), schizophrenia and Parkinson’s disease. Dopamine transporter plays a role in controlling the signaling and levels of Dopamine (2). DAT transports dopamine from the synapse and thus influencing the amount of dopamine in the nerve cells (2). Consequently, disruption of Dopamine …show more content…
DNA has negative charge due to the phosphate group in the DNA backbone. When the negatively charged DNA is exposed to an electric field in the gel, the DNA migrates towards the positive pole. The electrophoresis buffer contains ions that carry a current in the gel and provide a relatively constant pH in the gel. The distance of migration of the fragment is proportional to the size of the fragment. Therefore, gel electrophoresis in this practical was used to establish the presence of the DAT gene in the extracted DNA as well as determine the size of the DAT …show more content…
Lane 1, which contained water only used as the negative control, did not produce any band. Lane 2 with the contents of tube 2 with DAT VNTR PCR products from own buccal cells produced one band. Lane 3 loaded DAT VNTR PCR products from other partner buccal cells produced 2 bands as shown in fig 2. The lane 4 with the positive control DNA also produced one band. Figure 2: Gel Electrophoresis image showing the bands produced by controls and DAT PCR products of two individuals
The distance of migration of the bands for each sample was determined. The distance of migration of the bands obtained for the molecular size marker was used to draw a calibration curve using the log values of the size markers (fig. 3).
Figure 3: Calibration curve for the fragment size against distance of migration
From the calibration curve, the sizes of the DNA samples were determined: Distance migrated Log Fragment size
1 PCR tube 1 negative control 0 0 0
2 PCR tube 2 DNA sample 1 22.2 2.80 640bp
3 PCR tube 3 DNA sample 2 22.2
20.4 2.80
2.90 640bp
800bp
4 PCR tube 4 positive control DNA 22.2 2.80 640bp
The size of DNA in sample 1 is 0.
The size of DNA in sample 2 is
The dopamine transporter gene in human has had a strong association with certain neuropsychiatric disorders and behavioral conditions (1). This is because neurotransmission of dopamine underlies brain functions such as locomotion, cognition, behavior, and motivation (1). Interference with the production, regulation and signaling of dopamine and its neurotransmissions can cause neuropsychiatric diseases such as attention deficit/hyperactivity disorder (ADHD), schizophrenia and Parkinson’s disease. Dopamine transporter plays a role in controlling the signaling and levels of Dopamine (2). DAT transports dopamine from the synapse and thus influencing the amount of dopamine in the nerve cells (2). Consequently, disruption of Dopamine …show more content…
DNA has negative charge due to the phosphate group in the DNA backbone. When the negatively charged DNA is exposed to an electric field in the gel, the DNA migrates towards the positive pole. The electrophoresis buffer contains ions that carry a current in the gel and provide a relatively constant pH in the gel. The distance of migration of the fragment is proportional to the size of the fragment. Therefore, gel electrophoresis in this practical was used to establish the presence of the DAT gene in the extracted DNA as well as determine the size of the DAT …show more content…
Lane 1, which contained water only used as the negative control, did not produce any band. Lane 2 with the contents of tube 2 with DAT VNTR PCR products from own buccal cells produced one band. Lane 3 loaded DAT VNTR PCR products from other partner buccal cells produced 2 bands as shown in fig 2. The lane 4 with the positive control DNA also produced one band. Figure 2: Gel Electrophoresis image showing the bands produced by controls and DAT PCR products of two individuals
The distance of migration of the bands for each sample was determined. The distance of migration of the bands obtained for the molecular size marker was used to draw a calibration curve using the log values of the size markers (fig. 3).
Figure 3: Calibration curve for the fragment size against distance of migration
From the calibration curve, the sizes of the DNA samples were determined: Distance migrated Log Fragment size
1 PCR tube 1 negative control 0 0 0
2 PCR tube 2 DNA sample 1 22.2 2.80 640bp
3 PCR tube 3 DNA sample 2 22.2
20.4 2.80
2.90 640bp
800bp
4 PCR tube 4 positive control DNA 22.2 2.80 640bp
The size of DNA in sample 1 is 0.
The size of DNA in sample 2 is