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Constructing Restriction Maps with Partial Digestion Data in Cell Biology Labs

This document provides a comprehensive guide on constructing restriction maps based on partial digestion data using the BamH1 and EcoR1 restriction enzymes. It details the complete and partial digestion results revealing fragment sizes, which are integral for mapping DNA sequences. Included are methods for interpreting gel data and validation of constructed maps against observed digestion profiles. This activity is designed for students in BIO 330L Cell Biology Lab to enhance their understanding of molecular biology techniques and data analysis.

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Constructing Restriction Maps with Partial Digestion Data in Cell Biology Labs

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  1. More Partial Digestion Answer Key BIO 330L Cell Biology Lab Dr. Lin

  2. A)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 5, 6, 7, 8, 10, 11, 12, 13, 14, 18, 19 kb B)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 5, 6, 10, 11, 12, 13, 14, 16, 17, 18, 19 kb C)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5,  6, 7, 10, 11, 12, 13, 14, 17, 18, 19 kb D)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5,  6, 7, 10, 11, 12, 13, 16, 17, 18, 19 kb E)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5, 6, 7, 8, 10, 11, 12, 16, 17, 18, 19 kb

  3. (A) B B B B B 5 1 1 10 1 1 (B) B B B B B 5 1 10 1 1 1 (C) B B B B B 5 1 1 10 1 1 (D) B B B B B 1 5 10 1 1 1 (E) B B B B B 1 5 1 10 1 1

  4. F)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5, 6, 7, 8, 10, 11, 12, 13, 17, 18, 19 kb G)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5, 6, 7, 10, 11, 12, 15, 16, 17, 18, 19 kb H)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 5,  6, 7, 8, 10, 11, 15, 16, 17, 18, 19 kb I)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 4, 5,  6, 7, 8, 9, 10, 15, 16, 17, 18, 19 kb

  5. (F) B B B B B 1 5 1 10 1 1 (G) B B B B B 1 1 5 10 1 1 (H) B B B B B 1 1 1 5 10 1 (I) B B B B B 1 1 1 1 5 10

  6. J)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 16, 17, 18, 19 kb K)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 5, 6, 7, 8, 9, 10, 11, 12, 17, 18, 19 kb L)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 5,  6, 7, 8, 9, 10, 11, 12, 13, 18, 19 kb M)  Construct Restriction Map(s) base on the following Partial Digestion data using BamH1: Uncut: 19 kb BamH1 (Complete digestion) = 1, 5, 10 kb BamH1 (Partial digestion) =1, 2, 3, 4, 5, 6, 7, 8. 9, 10, 11, 12, 13, 14, 19 kb

  7. (J) B B B B B 1 1 1 5 1 10 (K) B B B B B 1 1 5 1 1 10 (L) B B B B B 5 1 1 1 1 10 (M) B B B B B 5 1 1 1 1 10

  8. Partial Digestion Answer Key (Enrichment activity #7) BIO 330L Cell Biology Lab

  9. Construct Restriction Map(s) base on the following Partial Digestion data using EcoR1: • Uncut: 21 kb; • EcoR1 (Complete digestion) = 2, 3, 4, 5, 7 kb; • EcoR1 (Partial digestion) = 2, 3, 4, 5, 7, 8, 9, 10, 12, 14, 17, 21 kb • The following are some of the possible Maps that fit the Gel data: • 1) 4E5E3E2E7 = 7E2E3E5E4 • 2) 4E3E5E2E7 = 7E2E5E3E4

  10. (A) E E E E 5 2 7 4 3 (A) E E E E 3 5 2 7 4 Uncut: 21 kb; EcoR1 (Complete digestion) = 2, 3, 4, 5, 7 kb; EcoR1 (Partial digestion) = 2, 3, 4, 5, 7, 8, 9, 10, 12, 14, 17, 21 kb (5 not at end: no16; 3 not at end: no18; 2 not at end: no 19)

  11. Construct Restriction Map(s) base on the following Partial Digestion data • using EcoR1: • Uncut: 21 kb; • EcoR1 (Complete digestion) = 2, 3, 4, 5, 7 kb; • EcoR1 (Partial digestion) = 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 21 kb The following are some of the possible Maps that fit the Gel data [Both of them are circular]: Map 1: 4E5E3E2E7E = 5E3E2E7E4E = 3E2E7E4E5E = 2E7E4E5E3E = 7E4E5E3E2E = 7E2E3E5E4E = 2E3E5E4E7E = 3E5E4E7E2E = 5E4E7E2E3E = 4E7E2E3E5E Map 2: 4E3E5E2E7E = 3E5E2E7E4E = 5E2E7E4E3E = 2E7E4E3E5E = 7E4E3E5E2E = 7E2E5E3E4E = 2E5E3E4E7E = 5E3E4E7E2E = 3E4E7E2E5E = 4E7E2E5E3E

  12. 4 E E 4 E E E 3 7 5 5 E E 7 3 7 3 E 5 E 2 E E 2 4 Accept: Both maps above fit gel data E E 2 E E Reject: Cannot get 10, 4 & 2 cannot be together (no 6 in partial digest) Uncut: 21 kb; EcoR1 (Complete digestion) = 2, 3, 4, 5, 7 kb; (every fragments can be at the end =>circular; [21-7=14, 21-5=16, 21-4=17, 21-3=18, 21-2=19]) EcoR1 (Partial digestion) = 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, [14, 16, 17, 18, 19,] 21 kb (4,7,2 need to be together to get 13; 4,2 cannot be together, no 6)

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