HMB265H1 Lecture Notes - Northern Blot

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raspberrysnake278

9 Feb 2012

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Dna from the organism extracted & cut into fragments with enzymes. Fragments separated into groups of fragments of different sizes (fractionated) by using electrophoresis. Fragments are blotted onto a piece of porous membrane where they maintain the same relative positions = southern blot -> probe for specific dna. Heat dna -> separate strands, the membrane is placed in a sol"n of the probe. The single-stranded probe will find & bind to its complementary dna sequence. On the blot, this binding concentrates the label in one spot -> relevant dna on gel is revealed. Find sets of genes -> use dna microarray -> genome wide probing ; array of dna fragments representing all the genes in a genome can be glued to the surface of a glass slide = microarray. Probes used are usually complex mixtures made by converting mrnas from one tissue into dna (cdna).

Related Questions

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.

7)DNA fragments are produced.

The order in which a DNA fingerprint is produced using gelelectrophoresis is:___,____,___,____,____,____,____,____

goldtrout506

Two "naked" (without histones or other proteins) double-stranded fragments of DNA are exactly the same length. At 89Â°C, fragment A has completely denatured, which means that the two strands have separated. At that temperature, fragment B is still double-stranded. How might these fragments differ, to result in different denaturation temperatures?

Restriction enzymes are used to construct restriction maps ofDNA. These are diagrams of specific DNA molecule that show thesites where the restriction enzymes cleave the DNA. To construct arestriction map, purified samples of the DNA are treated withrestriction enzymes, either alone or in combination, and then thereaction products are separated by agarose gel electrophoresis, atechnique that separates the DNA fragments based on size. (Thesmaller fragments travel more quickly and are found near the bottomof the gel, whereas the larger fragments are found near the top ofthe gel.)

Use the results of the agarose gel electrophoresis separation(above) to construct a restriction map for the sample of DNA.

apricothippopotamus885

HMB265H1 Lecture Notes - Northern Blot

HMB265H1 Full Course Notes

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