Name three techniques for SPECIFIC detection gene expression

Question 1

What is Northern Blotting ?

Answer 1

Technique which can be easily determined if we find out the amount of RNA transcribed in each tissue .

Question 2

Role

Answer 2

To detect the amount of RNA transcribed from gene x in each target tissue we need to

Question 3

Principle

Answer 3

1. Isolate total RNA from different target tissues
   2 Detect and quantify the specific RNA I.e transcribed from gene .
2. Blotting(RNA)
   Separated RNA molecules are transferred from the gel to the suitable nylon .
3. Hybridization and washing - Unbound probes are removed from washing

Hybridization - Need a probe that will specfically bind to target RNA molecules

X probe can be labelled complementary RNA or DNA sequence .

Nylon membrane is incubated with this probe molecules , probe specifically to their complementary target RNA molecules

Detection :

This depends on autodigrapy , flourecence colour or light forming reactions

Question 4

Main thing for gene expression

Answer 4

Determine when and where a particular gene is expressed

Aswell as to identify the presence of closely related species

Determine the size and abundance of RNA

Analysis of RNA processing

Question 5

What doe SITU hybridisation

Answer 5

Can be used to locate genes on isolated chromosomes and also for localising specific nuclei acids .

Question 6

There are two types of SITU hybridisation

Answer 6

In SiITU DNA hybridization : Precence and localisation of DNA sequence (gene/genome)

In SITU RNA hybridization : Presence and localisation of m-RNA sequences ( Level of gene expression /transcritome)

Question 7

Main thing

Answer 7

Is used to map and order genes and other DNA and RNA sequences to their location on chromosomes and within nuclei.

Question 8

Application

Answer 8

can be used to visually confirm sterility of fish tissues. This technique requires expertise in cryosectioning of tissues, in situ hybridisation, and fluorescent microscopy.

Question 9

S1 Nuclease Protection

Answer 9

Are extremely sensitive procedures for detection and quantitation of mRNA species in complex mixtures of total cellular RNA. …

A method describing S1 nuclease protection of target mRNA using either RNA or DNA probes is also included.