Laboratory exam

• parts of a micropipette

  hub

  setting ring

  release button

  pipette tip
• uses steam under pressure to kill harmful bacteria

  autoclave
• provides uniform temperature necessary for annealing, drying, and sterilizing

  oven
• spinning a variety of samples at high speed, pelleting of nucleic acids or proteins from solution

  micro-centrifuge
• determine sedimentation velocity, shape and mass of macromolecules, separation phases

  refrigerated centrifuge
• used for dna sequencing, cloning, dna quantification, patterns of gene expression

  thermocycler
• agitate particles in liquids, cell disruption

  sonicator
• removes moisture low enough to minimize chemical loss

  dehydrator
• analyze results of gel electrophoreses and blotting experiments, visualize stained nucleic acids and proteins

  gel doc imager
• measure chemical, biological reactions within the well of a microplate

  plate reader
• protection from exposure to biohazards

  biological safety cabinet (bsc)
• measure concentration of dna, rna, and protein

  nanodrop
• concentrating sample in batches

  concentrator
• technique for reproducing dna fragments

  dna cloning
• first stage of most genetic engineering experiments like pcr, dna sequencing, etc.

  dna cloning
• small, circular, double-stranded dna molecule that is distinct from a cell's chromosomal dna

  plasmid
• small piece of dna that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes

  cloning vector
• cleaves dna sequences at sequence-specific sites, producing dna fragments with a known sequence at each end

  restriction enzyme
• involves the exchange of genetic material either between multiple chromosomes or between different regions of the same chromosome

  dna recombination
• uptake of dna into bacterial, yeast or plant cells

  transformation
• amplification of a gene is coupled to the formation of product by the enzyme it encodes

  selective amplification
• process of determining the order of nucleotides bases in a piece of dna molecule

  sequencing
• reveals the genetic information that is carried in a dna segment

  sequencing
• sequencing reaction involves?


  replication of single-stranded template with the use of primer and dntp
• reaction mixture contains?


  taq dna polymerase

  all four dNTP's

  low concentration of ddNTP's
• monomers that are missing a hydroxyl group at the site at which another nucleotide usually attaches to form a chain

  ddNTP
• when incorporated on a growing complementary strand, it terminates the process of dna replication

  ddNTP
• dna is separated on the basis of

  size
• stops the synthesis of dna because of its lack of a free hydroxyl group needed for the replication of dna

  ddNTP
• What is the difference between the first steps of DNA sequencing to PCR in terms of the number of primers being used?

  pcr has 2 primers facing towards each other but sequencing has only one primer reading the sequence in one direction only
• What are the advantages of Next Generation Sequencing?

  ideal throughput per run

  quick and cost-effective
• separates dna molecules according to their size based on their rate of movement

  gel electrophoresis
• applications of gel electrophoresis


  dna fingerprinting

  analyze pcr

  analyze genes associated with disease

  dna profiling
• measure how much a chemical substance absorbs light by measuring the intensity of light that passes through a solution, each compound absorbs light over a certain range of wavelength

  spectrophotometry
• produce large amounts of identical copies sufficient for analysis

  pcr
• enzymes used to manufacture dna copies using a pre-existing dna molecules

  dna polymerase
• applications of pcr


  genotyping

  cloning

  sequencing
• specialized adaptations of pcr

  rt-pcr

  real time pcr

  multiplex pcr
• differentiate reverse transcriptase pcr and standard pcr

  pcr amplifies dna from a small amount of dna template, rt pcr uses reverse transcription to produce a dna template from an rna source than can then be amplified
• four steps for dna extraction

  lysis

  protease and rnase

  centrifugation

  precipitation