کی تمام اشاعتیں۔ UMAIR MASOOD . Abbottabad ، Pakistan
DNA synthesis by using Glycosyltransferases Micro Fluid Chip
DNA synthesis is a molecular biology process in which the DNA is created by using artificial method. There is different standard method for gene synthesis like: oligonucleotide synthesis, endonuclease-mediated assembly, annealing base connection of oligonucleotide. Oligonucleotide synthesis is methods of relatively short fragment of nucleotide with define sequence. The production of oligonucleotide the necessary bases adenine, thymine, guanine, and cytosine is added the synthesis is a computer control process that repeats several steps until full length product. Finally, the concentration of each oligonucleotide adjusted. The mixture of oligonucleotide is than provided gene synthesis lab. Oligo lab is assembled into the gene of your choice assembly by PCR (Polymerase chain reaction). DNA is made of nucleotides each nucleotide contains a phosphate group, a sugar (deoxyribose) and four nitrogenous bases adenine, guanine, cytosine, thymine. A deoxyribose sugar is the five carbon sugar molecules that help form the phosphate backbone of DNA. Deoxyribose sugar is a lacking of one oxygen atom that why called deoxyribose sugar. A deoxyribose sugar contains a “OH” group which play important role in our technique. Gene synthesis by using Glycosyltransferases Micro Fluid Chip method is simple and cost-effective method which DNA is synthesis by using syringe nozzle and micro fluid chip. Syringe consists of four types of nozzle and four types of syringes each nozzle is fill with different nucleotides (A,T, C,G). Nozzle drop it nucleotides drops on the micro fluid chip according to the sequence. The micro fluid chip carries short drops of nucleotides and Glycosyltransferases start adds DNA nucleotide according to the arrangement of drops or nucleotide.
HIV structure degradation and HIV cure:
When HIV first enters the human body or human blood stream the virus start circulates throughout the body. The protective surface of HIV is study with particular protein called gp120 and lip membrane envelop inside the capside that contain a viral RNA and viral enzyme.Gp120 protein on the virus is only able to bind with a cell surface called CD4-receptor.The second receptor protein ccr5 present on the surface of macrophages for the virus enter the cell by endocystosis and HIV cycle begin. From given Diagram virus Receptor attached with Lipid envelops if we can break the structure of Lipid envelops by using a enzyme called lipase virus could not be able to attach the surface of cell because the structure of the virus is degrade. The special type of lipase called pancreatic lipase which is working in 37°C and 7.0 PH can be used in this process
DNA fingerprinting and CRISPR cas9 system:
Leicester university geneticist Alec Jeffreys develops a technique called DNA fingerprinting in 1985 it allows the DNA sample from different people to be compared look for similarities and differences. It used the solving crime and can confirm if the people are related to each other like paternity testing. There is a section of chromosomes where instead of a gene consisting of a long sequence of bases, they are much shorter sequencing of three to five base that is repeated in many times these repeated sequences called short tandem repeats. If we can know the STR sequence of any person then we can design a gRNA.CRISPR to scan through DNA or find specific STR sequences of another person. If the CRISPR does not find the targeted STR sequence it does not bind to it its means that no fluorescence color appears under UV-light but its scan and find its target and this binding create a fluorescence signal its means that STR can occur in given DNA and both persons are related to each other with respect to any point that is father or son or suspect and victim. The DNA we can check is must be labeled with fluorescence color.
Mutation detection via CRISPRCas9 micro-fluid chip:
Author: Umair Masood
In biology mutation is a change in the nucleotide sequences of the DNA of an organism Mainly there are three types of mutation: point mutation, deletion and insertions. Once the mutation has been defined allele specific oligonucleotide hybridization, amplification, heteroduplex formation method referred to as a diagnostic method some advance technique like CRISPR cas9 system is using for selected mutagenesis. Using microfluid and CRISPR cas9 system we can detect a mutation. Let's say you have a DNA sample with fluorescent labeled from patient and you want to make sure that gene you are interested is in healthy gene. We can design a CRISPR to scan through DNA or find specific gene or mutation. The CRISPR scan the DNA if the CRISPR does not find targeted gene it does not bind to it its means that no fluorescence color appears under UV-light but its scan and find its target and this binding create a fluorescence signal its means that mutation can be occur in gene
Identify a gene that makes protein by using cell free protein synthesis system:
Author: Umair Masood
Living cell could genetically modify to perform a faction such as production of protein. However, these genetically modification often dispute with normal cellular function and result in mutation. Defect can be overcome through removing the bacterial membrane which leave the lysate that is perform both transcription and translation. The cell free-protein synthesis also known as invitro protein synthesis is the production of protein without uses of living cell. Gene is act as instruction to make protein. If we can isolate a gene and then apply a cell free protein synthesis system after synthesis the protein, run on gel-electrophoresis we can identify a gene on the basis of protein. Gel electrophoresis is a laboratory technique used to contrasting proteins according to molecular size and charge.
Protein sequence via nanopore sequencing machine:
Author: Umair Masood:
Protein is a biomolecule consist of one or more chain of amino acid residues. Protein have significance role in the structure function organization of the cell. Linear amino acid residues are called a polypeptide. Protein contain short and long amino acid polypeptide less than 20-30 residues. The building block of protein is amino acid which are small organic molecules consist of alpha carbon atom link to an amino group and carboxyl group hydrogen atom are variable components called side chain. All protein has primary secondary tertiary and quaternary structur.in the protein sequencing the major steps is first secondary tertiary and quaternary structure convert into the primer structure which is simpler as compare to other than primary structure is load into the nanopore sequencing machine. Nanopore sequencing machine work by monitoring changes to an electrical current as a nucleic acid are passed in a protein Nanopore, then signal is decoded and provide a particular DNA or RNA sequence our case nanopore sequencing machine work by monitoring change and electrical current as an amino acid which are passing through nanopore protein channel then signal is decoded and provide a particular amino acid sequence.
in-Vitro Site specific DNA Editing Via Restriction Enzyme:
Author: Umair Masood
full paper: https://www.academia.edu/44551733/In_Vitro_Site_specific_DNA_Editing_Via_Restriction_Enzyme
A few years ago, some researchers realized that they could use CRISPR cas9 to gene-editing tools. A part of DNA I like to change if I know the sequence of a gene. I can build a CRISPR that carry a matching code. one inside the cell CRISPR cas9 will scan the DNA until it finds that exact spots. when it does, it cut the DNA right there. now I have the broken gene I can inserted new sequence into the gap. Restriction digestion is a process in which DNA is cut at specific sites. Restriction enzyme bind and cleave double stranded DNA at specific site that main property of restriction enzyme used in DNA editing. Basically, the whole idea is depending upon two things: DNA denaturation and short fragment of DNA is bind to the template DNA and restriction enzyme cleave only double stranded DNA at specific site.
#design MSTN GENE Knockdown plasmid
MSTN basically limits the amount of myostatin.you can produce myostatin, myostatin limits how much mass your skeleton frame can hold so if you got no Mycostatin there is no limit to how big muscle you can get.