RNA Single-Round RNA Amplification and Biotin Labeling System

Technical Support Request Form

Low aRNA yields with experimental samples

• Input RNA does not contain poly(A) tracts >> RNA amplification with this kit requires the use of oligo dT primers. Do not substitute random or gene-specific primers with this kit.
  
  

Low yields with both experimental samples and control RNA

• Inadequate thawing and mixing of reagents >> Ensure reagents are completely thawed with no solids remaining. Gently mix and centrifuge reagents briefly.
  
• RNA sample vacuum centrifuged to dryness >> Monitor sample volume during vacuum centrifugation to avoid drying to completion. RNA is difficult to resuspend if dried completely.
  
• Inadequate elution of aRNA >> Elute nucleic acids from column filter purification systems with enough elution solution to completely saturate the filter. Dispense the elution buffer in the center of the column bed.
  
  

Average size of control aRNA is greater than 1kb but average size of experimental aRNA is less than 0.5 kb.

• Amount of input RNA is outside of recommended range (0.25 – 5 µg) >> If input RNA is below 250 ng, increase the amount of input RNA. If input RNA is above 5 µg, decrease the amount of input RNA.