bacteriologia

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Técnicas de biologia molecular da análise de genes e produtos gênicos únicos a abordagens em larga escala

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os mesmos genes, qual a diferença?

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Dogma central

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Localizando alvos
• Técnicas iniciais para evidenciar determinadas moléculas de interesse
• DNA / RNA métodos baseados em hibridização Generated by Foxit PDF Creator © Foxit Software http://www.foxitsoftware.com For evaluation only.

Hibridização desnaturação / renaturação de ácidos nucléicos

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Light absorption and temperature in DNA denaturation. (a) Melting of doubledstranded DNA can be monitored by the absorption of ultraviolet light at 260 nm. As regions of double-stranded DNA unpair, the absorption of light by those regions increases almost twofold. The temperature at which half the bases in a doublestranded DNA sample havedenatured is denoted Tm (for temperature of melting).
Light absorption by single-stranded DNA changes much less as the temperature is increased. (b) The Tm is a function of the G·C content of the DNA; the higher the
G·C percentage, the greater the Tm.

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Southern blot
• DNA é hibridizado com uma sonda – um fragmento de DNA de seqüência conhecida que contém algum tipo de marcação e pode ser detectado. • Mostra sequências complementares à sonda
• detecta pequenas quantidades (1 to 10 pg por banda) de uma sequência específica 1000 vezes mais