Random Hexamer Primers

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.

Applications

• 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