Random Hexamer Primer
Random Hexamer Primers are a mixture of oligonucleotides representing all possible sequence for that size. Random Primers can be used to prime synthesis in oligo-labeling similar to using hexamers and cDNA synthesis.
- Mixture of oligonucleotides -for RNA missing a polyA tail such as bacterial RNA
- Binds randomly -for RNA with strong secondary structure
Random Hexamer Primers consist of a mixture of oligonucleotides representing all possible hexamer sequences. Random Hexamer Primers are commonly used for priming single-stranded DNA or RNA for extension by DNA polymerases or reverse transcriptases.
During cDNA generation, random priming gives random coverage to all regions of the RNA to generate a cDNA pool containing various lengths of cDNA. Random priming is incapable of distinguishing between mRNA and other RNA species present in the reaction.
Primer Sequence: 5´ – d (NNNNNN) –3´ N = G, A, T or C
A mixture of random hexamer primers and oligo(dT) may improve the sensitivity of cDNA synthesis.
- DNA synthesis using Klenow fragment with DNA templates
- DNA probe synthesis for use in Northern and Southern blots, and in situ hybridization applications
- Partially degraded RNA samples
- RNA without poly(A) tail such as ribosomal RNAs
- RNA with strong secondary structure
- Target regions at 5´ end of a long messenger RNA transcript