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Molecular Biotechnology&Genomics

Information to learn off by heart

Information to learn off by heart

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20.01.2020 / 23.01.2020
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Illumina (Nucleotide detection, Read length, total bases/run, accuracy, cost(euro)/Mb)

  • Nucleotide detection: Reversible dye terminators
  • Read length: 100-300bp
  • total bases/run: 36 Gb/run
  • accuracy: 99%
  • cost(euro)/Mb: 0.1
  • Sequencing by synthesis

Pacific Biosystems 

  • Nucleotide detection: Phospho-linked fluorescent nucleotides
  • Read length: 100-14'000bp
  • total bases/run: 100 GB
  • accuracy: 85-95%
  • cost(euro)/Mb: 1.5
  • Sequencing by synthesis

Oxford Nanopore

  • Nucleotide detection: Change in membrane potential due to nucleotides
  • Read length: 4*103-105
  • total bases/run: 2.5-5
  • accuracy: 90%
  • cost(euro)/Mb: 0.5-1.0
  • Direct sequencing

3 Domninat markers + 3 auxotrophic markers 

(which ones have marker recycling and how?)

Auxotrophic

  • URA3, repairs uracil deficiency: negative selection with 5-FOA (marker recycling)
  • LYS2, repaires lysine deficienc: neg. sel with alpha-aminoadipate
  • MET15, repairs methionine deficiency: neg. sel with methyl mercury

Dominant markers (no marker recycling):

  • KanMX, resistance to G418
  • nat, resistance to nourseothricin
  • DsdA, resistance to D-serine

 

Restriction sites:

BsmBi, BsaI

BsmBi: 5' CGTCTC(N) 3'

BsaI: 5' GGTCTC(N) 3'

3 Type specific overhangs

CCCT

AACG

TATG

Stop codons (DNA sequence, 3)

TGA

TAG

TAA

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