DNA > Biotech > Flight_Manual > Northern blot

RNA

Minigel: RNA quality, quantity

Northern Gel

        Denaturing gel
        Running conditions
        UV Photograph

Transfer on membrane

        UV Illumination
        Fixation

Hybridization ---
 
        Salt
        Temperature
        Time

Probe

        Sequence
        Cross-hybridization
        Denaturation

Labeling

        Reaction efficiency
        Removal of unicorporated nucleotides
        Count

Washes (Geyger counter monitoring)
   
        Salt
        Temperature
        Time


Autoradiography
   
        Film
        Intensifying screens
        Low temperature
        Esposure time

Result interpretation (total/polyA+)

EXISTENCE / SIZE (expression)

    Cross hybridization

    Other isoform (5´, CDS, 3´)

    Other maturation stage

    Other gene (same family or not)


rRNA

QUANTITY

Pre-made filters - Example (Vitale et al., 2002)