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The document describes an experiment to determine the size of DNA fragments separated by agarose gel electrophoresis. A log plot of known DNA size standards versus migration distance was generated, allowing calculation of the size of unknown fragments. Six unknown bands were analyzed and calculated to be 19,331 bp, 9,823 bp, 6,651 bp, 4,382 bp, 2,230 bp, and 1,984 bp based on their migration distance. A figure illustrates the DNA ladder and placement of size labels for the unknown bands.

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17 views4 pages

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The document describes an experiment to determine the size of DNA fragments separated by agarose gel electrophoresis. A log plot of known DNA size standards versus migration distance was generated, allowing calculation of the size of unknown fragments. Six unknown bands were analyzed and calculated to be 19,331 bp, 9,823 bp, 6,651 bp, 4,382 bp, 2,230 bp, and 1,984 bp based on their migration distance. A figure illustrates the DNA ladder and placement of size labels for the unknown bands.

Uploaded by

om makwana
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Known Size Log of Size Measured Migration Log of

(bp) (mm) Migration


23130 4.36 19 1.27
9416 3.97 23 1.36
6557 3.82 26 1.41
4361 3.64 30 1.47
2322 3.37 39 1.59
2027 3.31 41 1.61
564 2.75
Table 1: Known sizes of DNA v/s distance measure of band migration in 1 % Agarose gel.

4.5
Log of DNA size standards

3.5

2.5 y = 5.6591x-1.13
2

1.5

1
1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8
Log of DNA fragment migration (in mm)

Figure 1. Log plot of DNA size standards (bp) and migration DNA fragments (in mm)

Calculations:

Band A migrated 19mm.

Log(19)=1.278

X=1.278

Y= 5.6591x^-1.13

Y=5.659 * 1.278 ^-1.13

Y= 4.28625699

10^4.28625699 = 19,331

Band A contains 19,331 base pairs.


Band B migrated 23 mm

Log (23) = 1.361727836

Y= 5.6591x^-1.13

Y=5.659 * 1.361727836 ^-1.13

Y = 3.992249155

10^3.992249155 = 9823

Band B contains 9823 base pairs.

Band C migrated 26 mm

Log (26) = 1.414973348

Y= 5.6591x^-1.13

Y=5.659 * 1.414973348 ^-1.13

Y = 3.822910762

10 ^ 3.822910762 = 6651

Band C contains 6651 base pairs.

Band D migrated 30 mm

Log (30) = 1.477121255

Y= 5.6591x^-1.13

Y=5.659 * 1.477121255 ^-1.13

Y = 3.641660431

10^3.641660431 = 4382

Band D contains 4382 base pairs.

Band E migrated 39 mm

Log (39) = 1.591064607

Y= 5.6591x^-1.13

Y=5.659 * 1.591064607 ^-1.13

Y= 3.34836245

10^3.34836245 = 2230

Bad E contains 2230 base pairs


Band F migrated 41 mm

Log (41) = 1.612783857

Y= 5.6591x^-1.13

Y=5.659 * 1.612783857 ^-1.13

Y= 3.297453078

10 ^ 3.297453078 = 1984

Band F contains 1984 base pairs.


Restriction endonucleases digests of Lambda
HindIII DNA

1 2 3 4

Size in base pairs

19,331
bp
9823
6651
4382
2230 bp
bp 1984

1 Commercial Lambda DNA +

2 Commercial Lambda DNA-

3 Digested Lambda HindIII+

4 Digested Lambda HindIII —

Figure 2: DNA fragment size labelled with their respected bands in a gel.

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