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Updated: Jun 17 2021

Molecular Biology Basics

  • Nuclear structures
    • Chromosomes
      • 46 in humans (23 pairs)
        • 22 pairs of autosomes, 1 pair of sex chromosomes
      • contains DNA and RNA
    • Deoxyribonucleic acid (DNA)
      • Thymine, adenine, guanine, and cytosine (TAGC)
        • adenine linked to thymine (A-T)
        • guanine linked to cytosine (G-C)
      • double stranded
      • strands linked together by phosphate groups
      • 2' hydrogen group
      • regulates cell division
      • mRNA is produced from DNA
        • an exon is portion of gene that codes for mRNA
        • exon is expressed
    • Gene
      • segment of DNA that contains the information needed to synthesize a protein
      • determines the unique biologic qualities of a cell
      • exon
        • coding information
      • intron
        • does not code for mRNA
    • Ribonucleic acid (RNA)
      • usually single stranded but can be double stranded
        • double stranded RNA found in some viruses
        • certain interactions between single stranded RNA in human cells can form double stranded RNA
      • has ribose sugar
      • Uracil, adenine, guanine, and cytosine (UAGC, no thymine)
      • less stable than DNA
      • 2' hydroxyl group
      • can be located in either the nucleus or cytoplasm
      • messenger RNA (mRNA)
        • translates DNA information into protein
      • ribosomal RNA (rRNA)
        • major part of ribosome, which helps synthesize a protein
      • transfer RNA (tRNA)
        • transfers amino acids to mRNA
  • Genetic Terms
    • Nucleotides
      • thymine, adenine, guanine, and cytosine
    • Codon
      • sequence of three nucleotides
      • each codon correlates to one of the 20 amino acids
      • linking of the amino acids create a protein
    • Gene promotor
      • regulatory portion of DNA that controls initiation of transcription
    • Gene enhancers
      • site on DNA that transcription factors bind to
      • regulate transcription
    • Transcription
      • DNA => mRNA
    • Translation
      • mRNA => protein
    • Haploid
      • Haploid is the amount of DNA in a human egg or sperm cell (half the amount of DNA in a normal cell)
    • Whole-genome sequencing 
      • Whole-genome sequencing (WGS) is the process of determining the entirety, or near entirety, of the DNA sequence of an organism's genome, including coding and non-coding DNA
    • Whole-exome sequencing 
      • Whole-exome sequencing (WES) allows sequencing of only the protein-coding regions of genes in a genome (known as the exome)
    • Genome-wide association sequencing 
      • Genome-wide association sequencing (GWAS) uses single-nucleotide polymorphisms (SNPs) to evaluate mutational variability among patients with a phenotype (or disease) of interest. 
  • Cell Cycle
    • Phases
      • G0
        • represents a "stable" phase
        • cells are diploid (2N) in the G0 and G1 phases
      • G1
        • initial growth phase
        • cells are diploid (2N) in the G0 and G1 phases
      • S
        • DNA replication/synthesis phase
        • cells become tetraploid (4N) at the end of S and for the entire G2 phases
      • G2
        • gap phase
        • cells become tetraploid (4N) at the end of S and for the entire G2 phases
      • M
        • mitosis phase
  • Apoptosis
    • Defined as programmed cell death
    • Requires a series of intracellular signaling events 
    • Different from cell lysis - where a cell releases its contents into the surrounding area
    • One hallmark of cancer is the cell's loss of apoptosis
  • Research techniques
    • Agarose gel electrophoresis
      • separates DNA based on size
      • DNA is negatively charged
      • gel exposed to electric field
      • smaller pieces moves through gel faster
    • Southern blotting
      • restriction enzymes cut up DNA
      • separate on agarose gel
      • identifies DNA sequence
    • Northern blotting
      • restriction enzymes cut up RNA
      • separate on agarose gel
      • identifies RNA sequence
    • Western blotting
      • SDS-PAGE gel
      • identifies protein
    • DNA ligation
      • combining different DNA fragments not found together naturally to create recombinant DNA
    • Plasmid vector
      • an extrachromosomal element, often circular, that can replicate and be transferred independently of the host chromosome
      • one example of the function of a plasmid is antibiotic resistance
      • can be introduced into bacteria in the process of transformation
    • Polymerase chain reaction (PCR)
      • DNA => DNA
      • a molecular biology tool used to generate many copies of a DNA sequence
      • uses "primers" specific to a segment of DNA
      • requires temperature-mediated enzyme DNA polymerase
    • Reverse transcription polymerase chain reaction (RT-PCR)
      • RNA => DNA
      • variant of polymerase chain reaction (PCR) used in molecular biology to generate many copies of a DNA sequence from fragments of RNA
      • RNA strand is first reverse transcribed into its DNA complement
      • amplification of the resulting DNA proceeds using polymerase chain reaction
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