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DNA Sequencing Exam Questions and Answers Graded A+ $7.99   Add to cart

Exam (elaborations)

DNA Sequencing Exam Questions and Answers Graded A+

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  • DNA Sequencing
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  • DNA Sequencing

DNA Sequencing Exam Questions and Answers Graded A+ what is first generation sequencing and what did it involve? - Answers sanger sequencing -> used radioactive dideoxynucleotide triphosphates (ddNTPs) as DNA chain terminators -> gel-based and ran on gel -> then moved to capillary pol...

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  • October 21, 2024
  • 8
  • 2024/2025
  • Exam (elaborations)
  • Questions & answers
  • DNA Sequencing
  • DNA Sequencing
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TutorJosh
DNA Sequencing Exam Questions and Answers Graded A+

what is first generation sequencing and what did it involve? - Answers sanger sequencing

-> used radioactive dideoxynucleotide triphosphates (ddNTPs) as DNA chain terminators

-> gel-based and ran on gel

-> then moved to capillary polymers that would do that in sequencing farms

what is the Phred score? - Answers quality score for each nucleotide

Q = -10log10 P

P = probability that the base is incorrect



log transformed probability that the base is incorrect



we want a score of >20!

What is P (phred score) derived from? - Answers 1. peak spacing (ratio of largest to smallest in window
of 7)

2. uncalled/called ratio in a window of 7 peaks

3. uncalled/called ratio in a window of 3 peaks

4. peak resolution

what does a Q score of 10 vs 20 mean?> - Answers 10: 1 in 10 probability wrong

20: 1 in 100 probability wrong

how did the human genome project work? - Answers make BAC libraries from human genome and then
sequence that

how was sequencing used clinically? - Answers used to look for SNPs

-> homozygous would have double the signal of heterozygous

how is 1st gen and 2nd gen (and 3rd gen) sequencing different? - Answers 1st gen:

gel-based or capillary sequencing

- long

, - template is not single molecule (PCR)

- $1/read

- 37M/10X haploid genome

-non-reversible terminators

- not clonal

- analogue detection of fluorophores



2nd gen:

massively parallel sequencing

- short (2*100 bases)

- template clonally derived from single molecule

- $0.00125/read

- 10K / 30X haploid

-reversible terminators

-clonal sequencing

- automatic detection and base calling



3rd gen

- single molecule reads

- very long reads

- low quality and reliability

how to construct 2nd gen sequencer library? - Answers DNA/cDNA

-> shear into Pool of Random fragments

-> end repair and ligate w/ adapters (known sequences)

-> PCR amplify

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