To Cleave Plasmid and Lambda Dna Using Restriction Enzyme
Autor: Jannisthomas • January 4, 2018 • 1,450 Words (6 Pages) • 705 Views
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E: Lambda DNA Cut With EcoRI
Distance Of Each Band(cm)
Log DNA Fragments Size
DNA Fragments Size(bp)
1.4
4.07
11749
18
3.87
7413
2.0
3.77
5888
2.1
3.72
5248
Lane E represents Lambda DNA cut with EcoRI hence it can be seen that it has multiple restriction sites along the DNA resulting in four bands.
F: Lambda DNA Cut With BglI
Distance Of Each Band(cm)
Log DNA Fragments Size
DNA Fragments Size(bp)
1.45
4.05
11210
1.60
3.97
9433
2.00
3.77
5953
2.30
3.62
4215
2.40
3.57
3757
3.20
3.18
1496
3.40
3.08
1189
3.50
3.03
1059
3.60
2.98
944
4.00
2.78
596
From Lane F it can be observed that BglI has many restriction sites along the DNA resulting in approximately 10 bands.
- Analysis of G, H & I
G: Plasmid Cut with Enzyme 1
Distance Of Each Band(cm)
Log DNA Fragments Size
DNA Fragments Size(bp)
2.20
3.67
4730
From lane G, it can be observed that the size of the plasmid is about 4730 bp.
H: Plasmid Cut with Enzyme 2
Distance Of Each Band(cm)
Log DNA Fragments Size
DNA Fragments Size(bp)
2.30
3.62
4215
4.20
2.68
473
From lane H, it can be concluded that since there are two bands therefore there are two restriction sites for Enzyme 2.
I: Plasmid Cut with Enzyme 1 & 2
Distance Of Each Band(cm)
Log DNA Fragments Size
DNA Fragments Size(bp)
2.50
3.52
3349
3.70
2.93
842
4.20
2.68
473
From lane I, it can be observed that the band of 473 bp is not being cut, this can be inferred from lane H as it also a band of 473bp. The other band from Lane H was cut into two pieces at 3349 bp and 842 bp.
The sum of the DNA fragments and the whole DNA is not equal these can be due to various systematic errors like the calibration of the scale used, random errors could have occurred while loading the sample, and measurement errors while loading the sample could have caused the difference in sum.
Experimental analysis and study questions – Lab 102
1. To which electrode does DNA migrate and why?
Ans: The DNA migrates towards the positive electrode since the DNA is negatively charged due to the presence of the phosphate groups.
2. Why has the discovery of restriction enzymes been so important?
Ans: Restriction enzymes allow the cleavage of DNA in a specific site; thus resulting in small fragments of DNA which can be used for various purposes like recombinant DNA technology, DNA sequencing, DNA fingerprinting
3. How are restriction enzymes named?
Ans: Restriction enzyme names consist of the first letter of genus, first 2 letters of the species; type of strain or substrain may follow as a capital letter; roman numeral designates one out of possibly several restriction enzymes produced by the same organism.
4. Briefly explain the function of restriction enzymes.
Ans: Restriction enzymes recognize and cleave DNA at specific sites resulting in DNA fragments whose ends are known and can be used for various applications like recombinant DNA technology.
Experimental
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