BIC472 Student Name: Darshan Trivedi
Partner’s Name: Manan
Assignment 2
POLYMERASE CHAIN REACTION
Total Marks: 12
1. (2 marks) In this experiment, you have amplified the D1S80 locus by PCR. Explain the advantages of using this locus to distinguish one person from another. Do you think you could use a coding gene for the same purpose? Clearly explain your answers.
D1S80 locus is placed on the short arm of the chromosome 1. This locus does not code for the arrangement for protein, yet it codes for a series of tandem repeats of 16 bp in human. Distinctive number of this allele has different number of repeats. These quantities of repeats are exceptional to every human. Primer
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Sr No.
Distance traveled by fragments in cm
Log molecular weight of each fragment
Molecular weight (bp)
1
11.7
2.7
501 bp
2
11.0
2.75
562 bp
Calculation:
Log molecular weight of the fragment of 1500 bp:=anti
Log (1500) = 3.18
Molecular weight of sample: (Genomic DNA Fragment # 2) = Antilog (2.75) = 562.
Number of repeats on Genomic DNA fragment #1: 113 bp + (n)(16 bp) + 32 bp = fragment size 113 bp + (n)(16 bp) + 32 bp = 501 bp (n)(16 bp) = 501 bp – 32 bp – 113 bp n = (356 bp / 16bp) n = 22.25 = 22 tandem repeats
The number of repeats on Genomic DNA fragment #1 is 22 Number of repeats on Genomic DNA fragment #2:
113 bp + (n)(16 bp) + 32 bp = fragment size 113 bp + (n)(16 bp) + 32 bp = 562 bp (n)(16 bp) = 562 bp – 32 bp – 113 bp n = (417 bp / 16bp) n = 26.06 = 26 tandem repeats
The number of repeats on Genomic DNA fragment #2 is 26
The number of repeat on Genomic DNA fragment #1 is 22 and on Genomic DNA fragment #2 is 26.
Genomic DNA is heterogenrous because it shows 2 fragments on 2% agarose gel which come from parents, mom and dad. Moreover, the tandem repeats(n) is within the standard limit (14-41) of heterogenic DNA. So, the sample is heteregenerous.
3. (2 marks) Ted just finished his PCR experiment and obtained the results in the
(PCR), which isolates small fragments of DNA that have a high degree of variability from
d. In 3.a (above) you mutated one letter. What role do you think the redundancy of the genetic code plays in this type of change?
For one of the monohybrid crosses you performed in this Investigation, describe how to use the phenotype ratios to determine
B) All humans are nearly identical genetically in coding sequences and have many proteins that are identical in structure and function. Nevertheless, each human has a unique DNA fingerprint. Explain this apparent contradiction.
The following results helped obtain the haplogroup that in which the sequence of mtDNA would identify. The PCR reaction worked, and this can be determined by looking at the agarose gel in figure 1. If the PCR reaction was successful, than a band should appear around 550bp. Individual AC displays a band around 550bp, this means the PCR reaction was successful. The band for individual AC, depicts a low concentration of product, because the band faint. After the purification process the concentration, A260/280 ratio, and A260/A230 ratio were determined by using the nanodrop. The concentration of mtDNA in the product was 60.9 ng/uL. The ratio for A260/280 was 1.79 and the ratio for A260/230 was 0.77. The A260 and 280 are a spectrometer measurement that measure absorbance at wavelengths of
Sample one has nearly identical amounts of dAMP and dTMP, and of dCMP and dGMP. thus this sample could be a duplex DNA, which implies that it's 2 strands of DNA that are complementary.
DNA molecules were unique to an individual with the exception of monozygotic twins. Due to
However, DNA fragments of 3 lab subjects didn’t show up on the gel. The allele
While we had performed sub cellular subscription function before the function of RNA isolation in 15 cell lines to deeply interrogate the human transcription. Now for the K562 lines we had perform the additional nuclear sub fractionation into the chromatin, nucleolus and the nucleoli. The RNA from each of these sub compartment were prepare in the form of replica and were separated from each other on the basis of the length into >200 nucleotides long and <200 nucleotides short. The part consisting of the long RNA were again been further fractionated into polyadenylated and in the form of non polyadenylated transcripts. There was a use of various number of complementary technologies were been use to characterize these fractions of RNA to their sequential order was made. Sequence rewards were been mapped and were been proceed through the use of a variety of tools also called as the software tools. We have been used to map the data to assemble those and quantify De novo elements. For the process of reproducibility the elements and the quantification were been further assessed between replicates and non parametric version of the irreproducible detection rate for the statistical test. The most part of the analysis with at least 90% irreproducible. Then the data which are been
Microsatellite and mitochondria DNA (mtDNA) genetic markers are often used in population genetic studies. Please detail the differences in their mode of inheritance, as well as what types of genetic information that these markers may provide.
A highly conserved gene will be used to identify a prokaryotic species isolated from the body. Fundamental lab techniques will be also explored and utilized, such as amplifying using PCR, cloning, and transforming the gene into a host cell. DNA electrophoresis and specific substrate plating will serve as analysis check points. The final product will be sequenced and compared to similar species to observe phylogenetic relationships.
Biological Background: It is scientifically proven that DNA (deoxyribonucleic acid) contains genetic material and highly specific to individual. DNA fingerprint can be tested by restriction fragment length polymorphism (RFLP) methods and Polymerase chain reaction (PCR) methods. Although RFLP is considered more accurate, due to the cost and requirement of longer period to complete, it is not commonly used. While only 1% of genetic materials are unique to individual, the short tandem repeats (STR) sequences called minisatellites can be used to distinguish all humans as it shows great variation between each person (What is DNA fingerprint? 2016).
The objective of the project is to determine if I carry the allele at the TAS2R38 locus. In order to find that out, DNA was extracted from cells within the mouth. The DNA sample was quantified, amplified by PCR, ran through gel electrophoresis, purified and sequenced. The results for my DNA was not able to be determined through sequencing. The DNA samples of the class recorded that 62% of the participants were tasters (carries the allele at the TAS2R38 locus) and 38% of the participants were non-tasters (does not carry the allele at the TAS2R38 locus).
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