Random Hexamer Labelling Flashcards
1
Q
What are hexamers?
A
Random, 6 nucleotide long oligos which have all possible variations.
2
Q
Why are hexamers 6 nucleotides long?
A
As restrictions sites, which are six nucleotides, repeat 1/40000 nucleotides it is thought that of six nucleotides long they will bind at least 1/40000 nucleotides.
3
Q
How can it be used to create labelled probes?
A
The dNTPs can be labeled with a radio label or biotin/ deoxygenin.
4
Q
Describe the steps in random hexamer labelling?
A
Denature the dsDNA at 95oC, mix with the hexamers allowing hybridisation, use a DNA polymerase with 5’-3’ polymerase but not exonuclease activity to extend the oligos with dNTPs which can be labelled.
5
Q
What is hexamer labelling used for?
A
To prime ssDNA for DNA polymerisation extension.
