Lab 10: DNA sequencing

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Lab 10: DNA sequencing

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Institution Lab 10: DNA Sequencing

Course Lab 10: DNA sequencing

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ddNTPs are labeled with fluorescent dyes in order to differentiate between the different nucleotides. True Who is the scientist for whom the dideoxy sequencing method is named Sanger What are the letters in this diagram called? haplogroups In DNA sequencing, how many different fluorescent dyes are used to label ddNTPs? 4 In a sequencing reaction, deoxyribonucleotides terminate a replicating segment of DNA, while dideoxy ribonucleotides allow it to continue. False What is the ratio of ddNTP to dNTP in the nucleotide mixture for the sequencing reaction? 1 ddNTP to 50 dNTP Which of the following statements is NOT true of both PCR & DNA sequencing? a. Both use ddNTPs to prematurely terminate DNA synthesis b. Both are based on in vitro DNA synthesis c. Both require a primer for initiationd. d. Both use DNA polymerase to catalyze the reaction Both use ddNTPs to prematurely terminate DNA synthesis How long ago did our human haplogroups diverge? Tens of thousands of years ago What are the clusters in this diagram called? haplogroups Which type of gel is used in DNA sequencing? polyacrylamide What is the name of the the output image of the sequencing process? electropherogram Which of the following is true about the Sanger method? a. The primer used in the reaction has a radioactive or fluorescent label on its 3' end for visualization. b. DNA sequencing is rarely done using automated systems. c. The dideoxynucleotides used in the reaction have a hydroxyl group at the 3' position. d. Each of the four reaction tubes contain only one of the four ddNTPs at a ratio of about 1 ddNTP/50 dNTP. e. The fragments in the four reaction tubes are separated on agarose gel. Each of the four reaction tubes contain only one of the four ddNTPs at a ratio of about 1 ddNTP/50 dNTP A mixture of ddNTPs and dNTPs is used in Sanger Sequencing. Which of the following statement is correct? dNTPs have a hydrogen in the 3' position instead of a hydroxyl group and thus stops DNA synthesis when added to a growing chain. Dideoxynucleotides are essential for: a. Primer design b. PCR c. SDS-PAGE d. DNA Sequencing e. Gel electrophoresis DNA Sequencing What is the importance of constructing a phylogenetic tree of human mitochondrial DNA?

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Lab 10: DNA sequencing
hh hh hh

ddNTPshharehhlabeledhhwithhhfluorescenthhdyeshhinhhorderhhtohhdifferentiatehhbetweenhhthehhdifferenthhnuc
leotides. True
Whohhishhthehhscientisthhforhhwhomhhthehhdideoxyhhsequencinghhmethodhhishhnamed Sanger
Whathharehhthehhlettershhinhhthishhdiagramhhcalled? haplogroups
InhhDNAhhsequencing,hhhowhhmanyhhdifferenthhfluorescenthhdyeshharehhusedhhtohhlabelhhddNTPs? 4
Inhhahhsequencinghhreaction,hhdeoxyribonucleotideshhterminatehhahhreplicatinghhsegmenthhofhhDNA,hhwh
ilehhdideoxyhhribonucleotideshhallowhhithhtohhcontinue. False
WhathhishhthehhratiohhofhhddNTPhhtohhdNTPhhinhhthehhnucleotidehhmixturehhforhhthehhsequencinghhreaction?
1hhddNTPhhtohh50hhdNTP
WhichhhofhhthehhfollowinghhstatementshhishhNOThhtruehhofhhbothhhPCRhh&hhDNAhhsequencing?

a.hhBothhhusehhddNTPshhtohhprematurelyhhterminatehhDNAhhsynthesishh
b.hhBothhharehhbasedhhonhhinhhvitrohhDNAhhsynthesis
c.hhBothhhrequirehhahhprimerhhforhhinitiationd.hh
d.hhBothhhusehhDNAhhpolymerasehhtohhcatalyzehhthehhreaction
BothhhusehhddNTPshhtohhprematurelyhhterminatehhDNAhhsynthesis
Howhhlonghhagohhdidhhourhhhumanhhhaplogroupshhdiverge? Tenshhofhhthousandshhofhhyearshhago
Whathharehhthehhclustershhinhhthishhdiagramhhcalled? haplogroups
WhichhhtypehhofhhgelhhishhusedhhinhhDNAhhsequencing? polyacrylamide
Whathhishhthehhnamehhofhhthehhthehhoutputhhimagehhofhhthehhsequencinghhprocess? electropherogram
WhichhhofhhthehhfollowinghhishhtruehhabouthhthehhSangerhhmethod?

a.hhThehhprimerhhusedhhinhhthehhreactionhhhashhahhradioactivehhorhhfluorescenthhlabelhhonhhitshh3'hhendhhforhhvi
sualization.
b.hhDNAhhsequencinghhishhrarelyhhdonehhusinghhautomatedhhsystems.
c.hhThehhdideoxynucleotideshhusedhhinhhthehhreactionhhhavehhahhhydroxylhhgrouphhathhthehh3'hhposition.
d.hhEachhhofhhthehhfourhhreactionhhtubeshhcontainhhonlyhhonehhofhhthehhfourhhddNTPshhathhahhratiohhofhhabouthh
1hhddNTP/50hhdNTP.hh
e.hhThehhfragmentshhinhhthehhfourhhreactionhhtubeshharehhseparatedhhonhhagarosehhgel.
EachhhofhhthehhfourhhreactionhhtubeshhcontainhhonlyhhonehhofhhthehhfourhhddNTPshhathhahhratiohhofhha
bouthh1hhddNTP/50hhdNTP
AhhmixturehhofhhddNTPshhandhhdNTPshhishhusedhhinhhSangerhhSequencing.hhWhichhhofhhthehhfollowinghhst
atementhhishhcorrect?
dNTPshhhavehhahhhydrogenhhinhhthehh3'hhpositionhhinsteadhhofhhahhhydroxylhhgrouphhandhhthushhstop
shhDNAhhsynthesishhwhenhhaddedhhtohhahhgrowinghhchain.
Dideoxynucleotideshharehhessentialhhfor:

a.hhPrimerhhdesign
b.hhPCR
c.hhSDS-PAGE
d.hhDNAhhSequencinghh
e.hhGelhhelectrophoresis DNAhhSequencing

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Uploaded on: October 21, 2024
Number of pages: 2
Written in: 2024/2025
Type: Exam (elaborations)
Contains: Questions & answers

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