GENETIC TEACHER

DNA Sequencing Method use DNA fragmentation for genomic DNA library preparation for sequencing and data analysis to study genes structure and mutations (Watch Related Video in #geneticteacher) #geneticteacher

RNA Sequencing Method use reverse transcription (RT-PCR) for cDNA library preparation for sequencing and data analysis to study gene expression and gene function and splicing patterns, Microarrays Method analyze large scale gene expression in many samples to detect changes of gene expression, SAGE Method discover new expressed genes or unknown gene transcriptions without require prior knowledge of sequences (Watch Related Video in #geneticteacher) #geneticteacher

DNA Sequencing Method use DNA fragmentation for genomic DNA library preparation for sequencing and data analysis to study genes structure and mutations, RNA Sequencing Method use reverse transcription (RT-PCR) for cDNA library preparation for sequencing and data analysis to study gene expression and gene function and splicing patterns, Microarrays Method analyze large scale gene expression in many samples to detect changes of gene expression, SAGE Method discover new expressed genes or unknown gene transcriptions without require prior knowledge of sequences (Watch Related Video in #geneticteacher) #geneticteacher

First Generation Sequencing include Maxam-Gilbert and Sanger Sequencing Methods, Second Generation Sequencing contain platforms of NGS Method (Watch Related Video in #geneticteacher) #geneticteacher

Next Generation Sequencing Method use parallel sequencing platforms to sequence multiple samples in parallel and sequence massively fragments at once (Watch Related Video in #geneticteacher) #geneticteacher

Sanger Sequencing Method is enzymatic sequencing method based on selective incorporation of chain-terminating nucleotides during DNA synthesis to manually or automatically sequence up to 1000 bp (Watch Related Video in #geneticteacher) #geneticteacher

Maxam-Gilbert Sequencing Method is chemical sequencing method based on nucleotide specific chemical modifications and subsequent DNA cleavage to manually sequence (200–300) bp (Watch Related Video in #geneticteacher) #geneticteacher

Maxam-Gilbert Sequencing Method is chemical sequencing method based on nucleotide specific chemical modifications and subsequent DNA cleavage to manually sequence (200–300) bp, Sanger Sequencing Method is enzymatic sequencing method based on selective incorporation of chain-terminating nucleotides during DNA synthesis to manually or automatically sequence up to 1000 bp, Next Generation Sequencing Method use parallel sequencing platforms to sequence multiple samples in parallel and sequence massively fragments at once, First Generation Sequencing include Maxam-Gilbert and Sanger Sequencing Methods, Second Generation Sequencing contain platforms of NGS Method (Watch Related Video in #geneticteacher) #geneticteacher

Next Generations Sequencing Method consider fast and scalable and accurate and effective sequencing method for large scale projects without require prior knowledge of genome to study genetic variations and transcriptional variations and genetic relationship, Next Generations Sequencing Method consider labor and costly method because it require computer capacity and experts in data analysis and statistics, Next Generations Sequencing Method use in large scale genomic studies and biological and medical studies and agriculture and environmental studies and pharmacogenomics studies and forensic studies (Watch Related Video in #geneticteacher) #geneticteacher

Next Generations Sequencing Method initiate by DNA or RNA extraction and library preparation in process of fragmentation by fragment DNA or RNA into shorter sequences (100-300) bp and ligate adaptors in both ends of fragments in process of indexing or barcoding and cluster generation by binding fragments to flow cell and undergo bridge amplification to form clusters for sequencing by using massively parallel sequencing platforms and data analysis by align output sequence to reference sequence (Watch Related Video in #geneticteacher) #geneticteacher

Next Generations Sequencing Method use in whole genome sequencing to sequence entire genome for complete DNA sequences and whole exome sequencing to sequence all exons in genome and target sequencing to sequence specific genes and RNA sequencing or transcriptome sequencing to sequence all RNA for gene expression profile and single cell sequencing to sequence single cell to understand biological processes and epigenetic sequencing to examine modifications that affect gene activity without altering DNA sequencing e.g. DNA methylation and proteomic sequencing to enable precise protein quantification and highly multiplex sequencing to process multiple samples in single run and chip sequencing to investigate protein DNA interaction (Watch Related Video in #geneticteacher) #geneticteacher

Next Generations Sequencing Method or High Throughout Sequencing Method is modern sequencing platforms sequence millions of nucleic acid fragments in parallel by break target nucleic acid into smaller pieces and attach adaptor to each fragment for sequencing (Watch Related Video in #geneticteacher) #geneticteacher

Next Generations Sequencing Method or High Throughout Sequencing Method is modern sequencing platforms sequence millions of nucleic acid fragments in parallel by break target nucleic acid into smaller pieces and attach adaptor to each fragment for sequencing, Next Generations Sequencing Method use in whole genome sequencing to sequence entire genome for complete DNA sequences and whole exome sequencing to sequence all exons in genome and target sequencing to sequence specific genes and RNA sequencing or transcriptome sequencing to sequence all RNA for gene expression profile and single cell sequencing to sequence single cell to understand biological processes and epigenetic sequencing to examine modifications that affect gene activity without altering DNA sequencing e.g. DNA methylation and proteomic sequencing to enable precise protein quantification and highly multiplex sequencing to process multiple samples in single run and chip sequencing to investigate protein DNA interaction, N...

Sanger Sequence Method consider gold standard for single genes or small DNA regions with base accuracy 99.99% up to 1000bp in single run without require complex bioinformatics tools and experts, Sanger Sequencing Method consider labor and costly and time consuming method and require relatively high quality of DNA samples leading to difficult to apply large scale sequences, Sanger Sequencing Method use in Mutation Studies and Epigenomic Studies and Medical Studies and Pharmagenomics Studies and Forensic Studies (Watch Related Video in #geneticteacher) #geneticteacher

Automatic Sanger Sequencing Method fluorescently label primer at 5’ end to amplify specific DNA regions and label each of ddNTPs with fluorescent dyes and run reaction in single lane in denature polyacrylamide gel electrophoresis or Capillary gel to visualize nucleotides peaks that reflect sequence of chain termination to compare output sequence with reference sequence (Watch Related Video in #geneticteacher) #geneticteacher

Manual Sanger Sequencing Method initiate by DNA extraction and denature DNA to obtain single DNA fragments and divide DNA sample into four tubes with primer and dNTP, and Taq DNA polymerase and one ddNTPs in each tubes to terminate DNA synthesis at specific base to create set of DNA fragments with different length after that separate amplified fragments in parallel lanes (A / G / C / T) in denature polyacrylamide gel electrophoresis and autoradiography gel in X-ray film or UV light to visualize length of amplified fragments and read bands from bottom to top according to direction of DNA synthesis (5’→3’) to compare sequence with reference sequence (Watch Related Video in #geneticteacher) #geneticteacher

Sanger Sequencing Method use single strand DNA for sequencing and primer to flank regions of target sequence and Taq DNA polymerase to synthesize new DNA strand by adding nucleotides to primer and dNTPs that add in synthesis process of DNA strand and ddNTPs to interrupt DNA strand synthesis as chain termination strand (Watch Related Video in #geneticteacher) #geneticteacher

Sanger Sequencing Method or Chain Termination Sequencing Method is first generation sequencing method use Dideoxy nucleotides (ddNTPs) that lack (3-OH) hydroxyl group and incorporate with growing DNA strand during PCR reaction as DNA chain terminators to generate fragments with ends ddATP / ddGTP / ddTTP / ddCTP, dNTPs contains (3-OH) hydroxyl group while ddNTPs contains hydrogen instead of (3-OH) hydroxyl group (Watch Related Video in #geneticteacher) #geneticteacher

Sanger Sequencing Method or Chain Termination Sequencing Method is first generation sequencing method use Dideoxy nucleotides (ddNTPs) that lack (3-OH) hydroxyl group and incorporate with growing DNA strand during PCR reaction as DNA chain terminators to generate fragments with ends ddATP / ddGTP / ddTTP / ddCTP, dNTPs contains (3-OH) hydroxyl group while ddNTPs contains hydrogen instead of (3-OH) hydroxyl group, Manual Sanger Sequencing Method initiate by DNA extraction and denature DNA to obtain single DNA fragments and divide DNA sample into four tubes with primer and dNTP, and Taq DNA polymerase and one ddNTPs in each tubes to terminate DNA synthesis at specific base to create set of DNA fragments with different length after that separate amplified fragments in parallel lanes (A / G / C / T) in denature polyacrylamide gel electrophoresis and autoradiography gel in X-ray film or UV light to visualize length of amplified fragments and read bands from bottom to top according to direc...

Maxam and Gilbert Sequencing Method use purified DNA directly in sequencing process to identify DNA modification without require PCR and consider accurate and suitable in DNA foot printing studies, Maxam and Gilbert Sequencing Method consider labor and costly and time consuming method and require hazardous chemicals as radioactive labels and Read short sequence length (50 to 500) bp, Maxam and Gilbert Sequencing Method use in DNA Structure Studies and DNA Modification Studies and DNA Interaction Studies and DNA Footpriting Studies and DNA Fingerprinting Studies (Watch Related Video in #geneticteacher) #geneticteacher

Maxam and Gilbert Sequencing Method initiate by DNA extraction and denature double strand DNA molecule to single fragments and radiolabel single fragments on 5’ ends using gamma 32P ATP After that cleave single fragments at specific positions using four chemicals reactions to generate breaks in specific nucleotides as follow: Tub- 1 contain dimethyl sulfate to break guanine nucleotide and Tube-2 contain dimethyl sulfate and formic acid to break purine nucleotides and Tube-3 contain hydrazine to break pyrimidine nucleotide and Tube-4 contain hydrazine and sodium chloride to break cytosine nucleotide and incubate four reactions tubes with piperidine to break modified strands and subject reactions side by side in denature polyacrylamide gel electrophoresis to differentiate fragments size according to radiolabels and autoradiography gel on X-ray film to read gel bands from bottom to top, Bands present in cytosine and pyrimidine nucleotides refer to cytosine nucleotide and bands present in...

Maxam and Gilbert Sequencing Method or Chemical Cleavage Sequencing Method is a first generation sequencing method cleave specific DNA molecule by certain chemicals and subsequent cleavage at specific bases as (G), (A+G), (C) and (C+T) which correspond to radiolabeled fragments (Watch Related Video in #geneticteacher) #geneticteacher

Maxam and Gilbert Sequencing Method or Chemical Cleavage Sequencing Method is a first generation sequencing method cleave specific DNA molecule by certain chemicals and subsequent cleavage at specific bases as (G), (A+G), (C) and (C+T) which correspond to radiolabeled fragments, Maxam and Gilbert Sequencing Method initiate by DNA extraction and denature double strand DNA molecule to single fragments and radiolabel single fragments on 5’ ends using gamma 32P ATP After that cleave single fragments at specific positions using four chemicals reactions to generate breaks in specific nucleotides as follow: Tub- 1 contain dimethyl sulfate to break guanine nucleotide and Tube-2 contain dimethyl sulfate and formic acid to break purine nucleotides and Tube-3 contain hydrazine to break pyrimidine nucleotide and Tube-4 contain hydrazine and sodium chloride to break cytosine nucleotide and incubate four reactions tubes with piperidine to break modified strands and subject reactions side by side i...