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3 Feb 2019

When chromatin from any eukaryote is digested with micrococcal nuclease (an endonuclease) and fractionated using electrophoresis, DNA fragments of approximately 200 base pairs in length are observed. Explain this result.

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Casey DurganLv2

4 Feb 2019

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NA fingerprinting uses a process called gel electrophoresis toseparate the fragments of DNA. Once the DNA fragments are sorted,the pattern of bands can be analyzed.

1)Gel Electrophoresis Procedure The smaller DNA fragments startto move away from the wells and the larger DNA fragments remaincloser to the wells.

2)An electric current is passed through the gel.

3) DNA fragments are treated with a dye.

4)A restriction endonuclease is added to the DNA.

5)Using micropipettes, the DNA samples are added to thewells.

6)DNA fingerprint is produced.

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You have been asked to clone the first enzyme of the leucine biosynthesis pathway from E. coli. To accomplish this objective, you have decided to purify the genomic DNA and treat with EcoRI (5â-G?AATTC-3â) restriction endonuclease. The genome size for E. coli is 6x106 bp (base pairs). After treatment with EcoRI with proper reaction conditions, you have found that the DNA was not digested by the restriction endonuclease EcoR1. However, when the same purified genomic DNA was treated with another restriction endonuclease BalI (5â-TGG?CCA-3â), the DNA was completely digested. The commercially purchased control E. coli DNA showed complete digestion under similar reaction conditions confirming that the EcoRI enzyme is functional and that the E. coli chromosome has multiple EcoRI restriction sites. What is the most probable cause for the E. coli DNA was not be digested by the EcoRI enzyme?

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13) You setup three PCR reactions using the same set of primers but with different DNA template samples (A, B, and C). You then perform agarose gel electrophoresis on the PCR reactions and stain the gel with ethidium bromide. When you look at the gel under UV light, one band is observed for each PCR reaction and all the bands are located at the same distance from the top of the gel. However, you notice that the band from Sample B is much darker than the bands for A and C. Which of the following is most likely true?

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B) The DNA fragments in sample B's PCR reaction are shorter than A and Câs.

C) The concentration of DNA in sample B's PCR reaction is less than C and A's.

D) The DNA fragments in sample B's PCR reaction are longer than A and Câs.

E) The DNA fragments in sample B's PCR reaction are less negatively charged than A and Câs.

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When chromatin from any eukaryote is digested with micrococcal nuclease (an endonuclease) and fractionated using electrophoresis, DNA fragments of approximately 200 base pairs in length are observed. Explain this result.

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