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