Which primer will anneal to this strand, and elongate during PCR? 5’ ATGCGGTTAC 3’ Group of answer choices 5’ GTAA 3’ 5’ ATGC 3’ 5’ TTAC 3’ 5’ GCAT 3’
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54.
Which primer will anneal to this strand, and elongate during PCR?
5’ ATGCGGTTAC 3’
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- 5. Show the separation pattern of the following DNA molecules on an agarose gel electrophoresis. 5 kbp 5 kbp 5 kb 5 kbp ----19. You run a PCR of the above HBB sequence using the primers you designed. You receive a vector from a company that you want to clone your HBB PCR product into. Below is the vector: BamHI 0.5 kb EcoRI 1.0 kb 0.3 kb -Bglll plasmid A 0.2 kb BamHI 0.4 kb HindIII 1.0 kb EcoRI You want to cut this vector with BamHI and EcoRI and insert your PCR product into this vector. After cutting the vector with both enzymes simultaneously you run a gel. i) On the gel below draw the bands you expect to see after you cut this vector with these enzymes at the same time (only use one gel lane - the other one is going to be empty). (6 points) Ladder 5kb 3kb 2.5kb 2kb 1.5kb 1kb 0.5kb 0.3kb 0.2kb ii) Circle the band that represents the backbone that you will insert the PCR product into. (2 points) c) Once you have inserted the PCR product into the plasmid backbone from part C, how many bp will the new vector be (the total of the backbone and the PCR product together). Answer should be 2 significant figures.…5' 3' O 60°C 5' O 95°C Primer 1 O 75°C ORF For your PCR reaction above, the primer set has a much higher AT (low GC) content than what would be considered normal. Due to this difference in AT content, the optimal annealing temperature for the primers is going to have to be altered from the "normal" temperature cycle: 95°C 60°C ⇒ 75°C Which of the temperatures will have to be altered to accommodate this difference the primer set? Primer 25¹ 3' 5' annealing with
- PCR primers Below is a 300 base pair fragment of DNA. The top strand is written in the 5' to 3' direction. The bottom strand is written 3' to 5'. There are also two primer sequences; both primers are written 5' to 3'. Note that we are displaying a double-stranded DNA fragment, but primers will only bind to one of the two displayed strands. 5' ACCOȚAGCTATATOCTATCOTGACOTATCOGCOCATTAAȚCGGGATCGAT 3 3' TGGCATCGATATACGATAGCACTGCATAGCCGCGTAATTAGCCCTAGCTẢ 5 50 5' AGCTCGCTAGCAGGAGAGATATCGCTCATAGCTCCGATCGATGCCGCTAA 3 100 3' TCGAGCGATCGTCCICTCTATAGCGAGTAICGAGGCTAGCTACGGCGATİ 5' 5' TATAGCTCTCTGCGGATATÇGCATẠTACCAAGGCCCTACGTATGTAGCTA 3 150 3' ATATČGAGAGACGCCTATAGCGTATATGGÍTCCGGGATGČATACATCGAŤ 5 5' TGCGȚATATÇGGAGAGTCCTGGATAT GGAGCTTGACTGCAGAGAGCTCGA 3 200 3' ACGCATATAGCCTCICAGGACCTATACCTCGAACÍGACGICTCTCGAGCİ 5' 5' TATGCGCTTAGGCCGTATATGCTTGGGGAAAGCTCTATGTATGCTATGTG 3 250 3' ATACGCGAATCCGGCATATACGAACCCCTITCGAĞATACATACG ẢTACAČ 5' 5' TGCATOTGCTATOCAACGTTC GGATTGCGȚAGCAGTAATAGCGCCGATTO 3' 300 3'…PCR primers Below is a 300 base pair fragment of DNA. The top strand is written in the 5' to 3' direction. The bottom strand is written 3' to 5'. There are also two primer sequences; both primers are written 5' to 3'. Note that we are displaying a double-stranded DNA fragment, but primers will only bind to one of the two displayed strands. 5' ACCGȚAGCTATATGCTATCGTGACGTATCGGCGCATTAAȚCGGGATCGAT 3 50 3' TGGCÁTCGATATACOATAGCACTOCATAGCCGCGTAATTÀGCCCTAGCTÀ 5' 5' AGCTÇGCTAGCAGGAGAGAȚATCGÇTCATAGCTCCGATCGATGCCGCTAA 3 3' TCGAGCG ATCGTCCTCTCTÁTAGCGAGTATCGAGÓCTAGCTACGGCGATİ 5' 100 5' TATAGCTCTÇTGCGGATATÇGCATATACCẠ AGGCCCTACGTATGTAGCTA 3 150 3' ATATČGAGAGACOCCTATAGCGTATATGGTTCCGGGATGČATACATCGAŤ 5' 5 TGCGTATATÇGGAGAGTCCTGGATATGGAGCTTGACTGCAGAGAGCTCGA 3 200 3' ACGCÁTATAGCCTCICAGGÁCCTATACCTCGAACTGACGTCTCTCGAGCT 5' 5' TATGCGCTTAGGCCGTATATGCTTGGGGAAAGCTCTATGTATGCTATGTG 3 3. ATACGCGAATCCGGCATATACGAACCCCTÍTCGAGATACATACGATACAC 5' 250 5' TGCATGTGCTATGCAACGTTCOGATTGCGȚAGCAGTAATAGCGCCGATTG 3 300 3'…For the following short sequence of double stranded DNA, design primers (just ~ 3-4 bases) and show 2 copy cycles of PCR (refer to figure 13.25) for the amplification of this sequence of DNA (so that you have 4 double stranded DNA). 5’- GGTATTGGCTACTTACTGGCATCG- 3’ 3’- CCATAACCGATGAATGACCGTAGC- 5’
- Choices: Origin of replication Bubble SSBP RPA Sliding Clamp PCNA DNA Pol III Pol ε Pol δ DNA Pol I RNAse H Flap 1 DNA gyrase Pol α DNA helicase Primase Single chromosome Multiple points in chromosomes DNA ligase Not applicable 12.Proof reading in eukaryotic DNA material 13.Removal of eukaryotic primer 14.Adds more nucleotides in the lagging strand of eukaryotes 15.Holds the processive enzyme in eukaryotes 16.Holds the processive enzyme in prokaryotesImage 1. Which 2 primers from the choices provided would work to amplify the DNA sequence given below ? 5’ACTGAGTCCATGCGATCATGACTAT 3’ 3’TGACTCAGGTACGCTAGTACTGATA 5’ this is a hypothetical example. In a real experiment Choose 5’ TGAC 3’ 5’ CTAT 3’ 5’ ACTG 3’ 5’ ATAG 3’ Image2. the template strand?The results of a gel-based sequencing experiment are shown below. What is the sequence, written Only include nucleotides (no spaces or numbers )25. The restriction enzymes Kpnl and Acc651 recognize and cleave the same 6-bp sequence. You have a plasmid and a linear DNA strand that both contain a Kpnl and Acc651 sequence in the same orientation as shown below. You digest both DNA pieces with both enzymes and then attempt to ligate the sticky ends, followed by treatment with DNA ligase. What will happen? 5' GGTACC3' 5' G G T ACC 3' 3' CCATGG 5' y CCATGGS Kpnl Acс651 A) You will produce sticky ends but the two types of ends will not ligate. Instead, you may produce a small amount of religated plasmid where the digested plasmid sequence re-inserts. B) You will produce a recombinant plasmid in which the linear DNA strand is ligated in between the two sites, suitable for cloning. C) You will produce blunt ends that will not ligate because the two restriction enzymes will both operate on both of the sites. D) All the DNA will be completely digested as if you had applied a general DNAse enzyme.
- 1. Why do we need to check the isolated DNA for its quantity and quality? 2. The purity of DNA sample is below 1.8 A260/A280 so, where did the proteincontamination come from? Note: Cite the in-text citation and references27. Given the following plasmid map, list how many fragments would exist and their bp length when treated with Hindll and Nde1 Dail 91 Ben BI 51 EstAPI 179 BemBI 2683 Ndel 183 Kasl - Narl - Sfol 235 Bgll 245 Fspl 256 Pul 276 EcoO1091 2674 Aatll - Zral 2617 BiM 2542 Sspl 2501 Prull 306 Benrl 364 Acll 2297 BæAl 387 Apol - EeoRI 396 Xmnl 2294 lacza haall - Sacl - Ecos3KI 402 Accbs1 - Kpnl 400 Awal- BsoN - Smal- TapM I- Ymal 412 Beal 2215 MCS Scal 2177 haml 417 Xhal 0 Pvul 2066 Accl - Hinl - Sall 29 pl. Alau a sb 434 Avall 2059 PUC19 2,686 bp BerDI 1985 Sphi 441 Hindi 447 Acll 1924. Fspl 1919 Prull 628 Avall 1837 Til 641 EsaXI 659 me AllI 1822 Rgll 1813 Bpal 1784 BSPOI - Sapl 683 BsrFl 1779 Bsal 1766 Thl 781 AII - Peil 806 BerDI 1753 Dal 908 Bmrl 1744 ori Aldl 1694 BkiM 1015 BseYl 1110 AlwNI 1217 BeeN 1292 dy8. You perform a PCR experiment using the template and primers shown below. What size is the amplicon (aka the PCR product) in bp? Template Q8: 5' TCGGC CGTAC TACGA TCGAT TGCGT ACGCA ATCGC -3' ¦¦¦¦¦ ¦¦¦¦¦ ¦¦¦¦¦ ¦|||| |||||||||| 3' AGCCG GCATG ATGCT AGCTA ACGCA TGCGT TAGCG -5' Primers: 5'-CGTAC-3' & 5'-TTGCG-3'