to which nucleotides might you design your guide RNA? Why? d) RNA Seq is used to determine off-target effects of Cas9 cleavage. Why is this an appropriate tool to determine these effects? e) Data on the following page is RNA-Seq data about indels that were detected away from the target site. What does this show about the effectiveness of CRISPR? Please also comment about the impact of these off-targets impacts on the utility of CRISPR in human medicine. ATACAAAGGAT CAGAAT CC CNG G . On-target chr465163142 T C FF chr1:199581904 chr22:47094773 chr487529947 chr9:109570965 chr7:132754767 chr2203801137 chrX:155356680 chrX:155484568 chr22:45689924 chr3:107039143 chr9:122015027 chr7:147113997 chrX:12851965 chrX:102677950 chr560703887 chr5:12506322 chr4:116565656 chr10:10467604 TC chrX:144268686 chr7:146700912 CA chr2:30757626 chr2:13854278 chr15:73766400 chrX:13727578 chr7:143114275 chr2:14708587 chr823569322 chr2216830092 chr20:17814888 T- chr8:135042874 C. TA. T . . ........... <.........ooooo.. C C ............ G G T с ....... G G G сст сст ···········.o. ... TT G C TC ..........000. G TC . ......... ...A CT ....
Molecular Techniques
Molecular techniques are methods employed in molecular biology, genetics, biochemistry, and biophysics to manipulate and analyze nucleic acids (deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)), protein, and lipids. Techniques in molecular biology are employed to investigate the molecular basis for biological activity. These techniques are used to analyze cellular properties, structures, and chemical reactions, with a focus on how certain molecules regulate cellular reactions and growth.
DNA Fingerprinting and Gel Electrophoresis
The genetic makeup of living organisms is shown by a technique known as DNA fingerprinting. The difference is the satellite region of DNA is shown by this process. Alex Jeffreys has invented the process of DNA fingerprinting in 1985. Any biological samples such as blood, hair, saliva, semen can be used for DNA fingerprinting. DNA fingerprinting is also known as DNA profiling or molecular fingerprinting.
Molecular Markers
A known DNA sequence or gene sequence is present on a chromosome, and it is associated with a specific trait or character. It is mainly used as a genetic marker of the molecular marker. The first genetic map was done in a fruit fly, using genes as the first marker. In two categories, molecular markers are classified, classical marker and a DNA marker. A molecular marker is also known as a genetic marker.
DNA Sequencing
The most important feature of DNA (deoxyribonucleic acid) molecules are nucleotide sequences and the identification of genes and their activities. This the reason why scientists have been working to determine the sequences of pieces of DNA covered under the genomic field. The primary objective of the Human Genome Project was to determine the nucleotide sequence of the entire human nuclear genome. DNA sequencing selectively eliminates the introns leading to only exome sequencing that allows proteins coding.



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