1107LWk11GIRameshA

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1107L

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Biology

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Jan 9, 2024

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BIOL 1107L Gel Interpretation 1) You have to identify two unknown plasmids using Restriction Digest analysis. You miss lab when your group performs gel electrophoresis of your digests of plasmids 1 and 2 using different combinations of the enzymes Eco RI, Bam HI, and Hind III. They send you a picture of the gel showing their results (above), but they forget to tell you which lane corresponds to which digest. All you know is that lanes 1 and 10 are the molecular weight markers (DNA Ladder) with bands at 8000, 5000, 3000,1000, 500, and 100 base pairs (bp). Use the information from the gel and restriction digest maps to answer the questions below. (1 pt each) a. Which lane would represent the products of a digest of Plasmid 1 with EcoRI? The answer would be well 3 because Plasmid 1 with EcoRI would have 2 fragments, meaning that there would be two bands. This means we can dwindle down the options to lane 3 and 6. The bands would be around 1900 and 3100. Since lane 6 sits directly MWM 8,000 bp 5,000 bp 3,000 bp 1,000 bp 500 bp 100 bp MWM 8,000 bp 5,000 bp 3,000 bp 1,000 bp 500 bp 100 bp

BIOL 1107L Gel Interpretation at 3000 instead of slightly above like lane 3 (which indicates a length slightly above 3000), lane 3 is more likely to represents the products. b. Which lane would represent the products of a digest of Plasmid 2 with Bam HI and EcoRI? Lane 4 because there would be 3 bands, meaning we are choosing between 4 and 9. Since the longest band would be 2850, 4 matches better than 6 which has a band above the 3000 mark. c. Identify which plasmid was cut with Bam HI and resulted in Lane 5. Plasmid 1 with Bam Hi because there is only one location where the restriction enzyme would cut the DNA. 2) Would you categorize gel electrophoresis as a qualitative or quantitative analysis? Explain your reasoning. (2 pts) Gel electrophoresis is qualitative analysis because although it does show the length of the strands due to the speed of their movement, it does not have an exact number value assigned to it. Although there is a scale present to analyze the length of the strand, but the number is not exact and therefore it cannot be quantitative. 3) Using the information in the table of fragments calculated from the Restriction Enzymes Worksheet, analyze the results of the gel that was imaged in lab. Based on your sample, determine which primer set corresponds to your letter code.

BIOL 1107L Gel Interpretation Label of Unknown Primer Set: U G A Record all the bands observed for your sample wells of the gel results: (2 pts) Based on the results of bands observed, what do you think is the unknown primer set? (1 pt) Identity of Unknown Primer Set: 1 2 3 Explain the rationale for your selection. (2 pts) Using the DNA ladder, we can see the total size of the PCR strand uncut and then the stands that were results of restriction digest. By comparing the size of the stands to the amplified region found on the restriction enzymes worksheet to our gel electrophoresis, we can see that the strand size matches the results of Primer 2 approximately. Sample Name Lane # Bands Observed (bp) Control (uncut PCR product) 5 4400 EcoRI Digest 7 3100, 1200 HindIII Digest 6 100, 2000, 2100

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