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1. Calculate the size of the resulting fragments as they will occur after digestion and write the sizes on the maps below. Note that linear DNA has a total size of 48 502 bp (see figure 3A).

1. Calculate the size of the resulting fragments as they will occur after digestion and write the sizes on the maps below. Note that linear DNA has a total size of 48 502 bp (see figure 3A).

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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23:15 O O 0• 4G+ ll 8% Restriction enzyme tutorial 2021_1_-3 - Saved A DNA (bp) A 10,000 20,000 30,000 40,000 48,502 Bamtl 5505 22,346 27,972 34,499 41,732 (bp) ECOBI 21,226 26,104 31,747 39,168 44,972 (bp) D Hindu 23,130 27,479 25,157 36,895 37,584 44,141 37,459 (bp) Figure 3: Restrictrion site map showing the following: A) linear DNA that is not cut as reference B) DNA cut with BamHI, C) DNA cut with ECORI, D) DNA cut with Hindlll 1. Calculate the size of the resulting fragments as they will occur after digestion and write the sizes on the maps below. Note that linear DNA has a total size of 48 502 bp (see figure 3A). В BamHI 5505 22,346 27,972 34,499 41,732 (bp) C ECORI 21,226 26,104 31,747 39,168 44,972 (bp) D Hindll 23,130 27,479 36,895 37,584 37,459 44,141 (bp) 25,157 2. How many fragments would you expect to see for each of the maps indicated in question 1 above? Мар В: Мар С: Map D: 3 3. Use the sequence indicated in figure 1 (also refer to figure 2 and 3) and indicate how many times does the sequence GAATTC occur in the A DNA sequence? What about B IU A II || ||
Transcribed Image Text:23:15 O O 0• 4G+ ll 8% Restriction enzyme tutorial 2021_1_-3 - Saved A DNA (bp) A 10,000 20,000 30,000 40,000 48,502 Bamtl 5505 22,346 27,972 34,499 41,732 (bp) ECOBI 21,226 26,104 31,747 39,168 44,972 (bp) D Hindu 23,130 27,479 25,157 36,895 37,584 44,141 37,459 (bp) Figure 3: Restrictrion site map showing the following: A) linear DNA that is not cut as reference B) DNA cut with BamHI, C) DNA cut with ECORI, D) DNA cut with Hindlll 1. Calculate the size of the resulting fragments as they will occur after digestion and write the sizes on the maps below. Note that linear DNA has a total size of 48 502 bp (see figure 3A). В BamHI 5505 22,346 27,972 34,499 41,732 (bp) C ECORI 21,226 26,104 31,747 39,168 44,972 (bp) D Hindll 23,130 27,479 36,895 37,584 37,459 44,141 (bp) 25,157 2. How many fragments would you expect to see for each of the maps indicated in question 1 above? Мар В: Мар С: Map D: 3 3. Use the sequence indicated in figure 1 (also refer to figure 2 and 3) and indicate how many times does the sequence GAATTC occur in the A DNA sequence? What about B IU A II || ||
Section A: Linear DNA Common restriction enzymes include: EcoRI, Hindll and BamHl and their sequences are as follows, with the cut site indicated by the arrow (figure 1). Please note that A DNA refers to linear DNA in this tutorial. HindIII 5'..A AGCT.3' 3'....TTCGA A...5' EcoRI 5'..G AATTC...3 BamHI 5'....G GATCC...3' 3' .....CTTAA G..5 3'...CCTAG G...5' Figure 1: Restriction sites of restriction enzymes. When DNA is cut with restriction enzymes, the fragments can be seen on an agarose gel (see figure 2). Base Pairs 21220 25.000 10.000 8,000 6.000 5,000 6557 4361 1641 7233 4,000 3,000 2,500 7421 2.000 5804 E643 1,500 1.000 4878 750 -564 S00 E 3530 125 250 A cut with EcORI A cut with Hindil A cut with BamHI A C D Figure 2: DNA fragments on an agarose gel showing the following: A) a molecular ladder, B) DNA cut with EcoB, C) DNA cut with Hipd, D) DNA cut with Bam The above figure shows the size of each of the fragments/bands produced when A DNA is cut with each of these restriction enzymes. The sizes were determined by comparison to a molecular ladder (figure 2A) which has bands of known sizes when it is separated by electrophoresis at the same time as the digested A DNA. Restriction sites of Lambda (A) DNA - in base pairs (bp) The sites at which each of the 3 different enzymes will cut the same strand of lambda DNA are shown in the maps (see figure 3 B-D), each vertical line on the map is where the respective enzymes will cut. ||
Transcribed Image Text:Section A: Linear DNA Common restriction enzymes include: EcoRI, Hindll and BamHl and their sequences are as follows, with the cut site indicated by the arrow (figure 1). Please note that A DNA refers to linear DNA in this tutorial. HindIII 5'..A AGCT.3' 3'....TTCGA A...5' EcoRI 5'..G AATTC...3 BamHI 5'....G GATCC...3' 3' .....CTTAA G..5 3'...CCTAG G...5' Figure 1: Restriction sites of restriction enzymes. When DNA is cut with restriction enzymes, the fragments can be seen on an agarose gel (see figure 2). Base Pairs 21220 25.000 10.000 8,000 6.000 5,000 6557 4361 1641 7233 4,000 3,000 2,500 7421 2.000 5804 E643 1,500 1.000 4878 750 -564 S00 E 3530 125 250 A cut with EcORI A cut with Hindil A cut with BamHI A C D Figure 2: DNA fragments on an agarose gel showing the following: A) a molecular ladder, B) DNA cut with EcoB, C) DNA cut with Hipd, D) DNA cut with Bam The above figure shows the size of each of the fragments/bands produced when A DNA is cut with each of these restriction enzymes. The sizes were determined by comparison to a molecular ladder (figure 2A) which has bands of known sizes when it is separated by electrophoresis at the same time as the digested A DNA. Restriction sites of Lambda (A) DNA - in base pairs (bp) The sites at which each of the 3 different enzymes will cut the same strand of lambda DNA are shown in the maps (see figure 3 B-D), each vertical line on the map is where the respective enzymes will cut. ||
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