Aula de técnicas e marcadores moleculares

1. TECNICAS DE BIOLOGÍA
MOLECULAR

2. ENDONUCLEASAS
O ENZIMAS DE RESTRICCIÓN
 Generan cortes internos en la molécula de
ADN
 Pueden generar extremos romos o
cohesivos

3. Enzimas de Restricción

4. Extremos cohesivos

6. Extremos romos

8. Clonación y uso de enzimas

11. Separación de fragmentos
 Geles de Agarosa
 Geles de
poliacrilamida

13. CLONACIÓN

15. PCR (reacción en cadena de
polimerasa) Componentes de la
reacción
 Polimerasa
 Primers
 dNTP’s
 Cloruro de magnesio
 Agua
 Buffer

29. SOUTHERN BLOT

30. 1) Blood sample. 2) DNA is extracted from blood cells. 3) DNA is cut into
fragments by a restriction enzyme. 4) The DNA fragments are separated into
bands during electrophoresis in an agarose gel. 5) The DNA band pattern in the gel
is transferred to a nylon membrane. 6) Radioactive DNA probe, designed to
recognise specific DNA sequences, is prepared. 7) Probe DNA binds to the specific
DNA sequences on the membrane. 8) Excess DNA is washed off, but the DNA
pattern is invisible at this stage. 9) X-ray film is placed next to the membrane &
exposed to the radioactive DNA. 10) The developed X-ray shows the pattern of DNA
bands

36. SECUENCIACIÓN

44. VNTR

49. DNA fingerprinting in Argentina to
establish kinship of the
'disappeared', the children, often
born in captivity, of parents
kidnapped and assassinated by the
former military regime there. Here a
geneticist uses a multi-channel
pipette to prepare samples for the
process. A bank of blood samples,
taken from all the potential relatives,
provides the DNA for comparison
with that from the child in question.
Fragments of DNA are seperated
into bands by electrophoresis in an
agarose gel. Every human being
produces a unique pattern of DNA
bands, hence its potential as a kind
of genetic

50. CLONACIÓN EN ANIMALES

51. COPY-CAT (EE.UU 2002)

52. PAMPA (Argentina 2002)

53. PROMETEA (Italia 2003)

54. Ian Wilmut and Dolly. In 1996, British embryologist
Professor Ian Wilmut created, the world's first sheep
cloned from an adult sheep cell. The research was
conducted at the Roslin Institute in Edinburgh,
Scotland.

55. Marcadores moleculares
Y sus aplicaciones

56. Tipos de Marcadores
 Directos o sea dentro del gen
(intragénicos)
 Indirectos usando a otro gen o marcador
cercano en el cromosoma para poder
identificar sus diferencias entre individuos

57. RFLP’s
 Pueden hacerse por Southern blot
 Pueden hacerse por PCR

58. Utilidad
 Detección de individuos portadores de un
gen recesivo.
 Test de paternidad o trazabilidad.
 Selección asistida por marcadores,
 Salud publica,
 Virología, etc

59. Por
Southern
blot

61. Por PCR y luego restricción

63. VNTRs o microsatélites
 Pueden hacerse por Southern blot
 Por PCR y geles de poliacrilamida o
agarosa
 Por SSCP

71. Cepas diferentes de Salmonellas