Synthetic Carrier RNA (1 mg/ml or 10 mg/ml solution)
Product Description
highQu Synthetic Carrier RNA is designed to be used in all kind nucleic acid purification and precipitation procedures
as a carrier and co-precipitant of nucleic acids. It is especially useful to increase the amount of RNA or DNA pellet
in low concentrated solutions, in such procedures, as viral RNA extraction from human specimen samples.
In contrast to commonly used carrier RNAs such as tRNA, yeast RNA, or sonicated salmon sperm DNA,
the Synthetic Carrier RNA is free from animal or yeast RNA contamination. Coprecipitated RNA
and DNA can be directly used for all common downstream applications, such as PCR or RT-PCR,
as well as highly sensitive qPCR.
The use of carrier RNAs for coprecipitation of nucleic acids may interfere with spectrophotometrical concentration measurements.
The presence of carrier RNAs in the RNA or DNA solution may have some influence on certain enzymatic reactions
performed by such enzymes that act on all nucleic acid molecules, for example T4 Polynucleotide Kinase or Terminal DNA Transferase.
Useful tips how to use coprecipitates such as carrier RNA for purification or concentration of PCR products
and other low abundance DNA molecules without the use of PCR purification kits:
Use an appropriate coprecipitant concentration: Too little coprecipitant may result in poor recovery of PCR fragment,
while too much can lead to increased background noise when performing concentration measurements.
Use Synthetic Carrier RNA in DNA, PCR reaction product or RNA solutions during alcohol precipitation step.
To maximize the yield of nucleic acids, before adding salt (Sodium acetate) and Ethanol or Isopropanol,
first add Synthetic Carrier RNA and mix it well with DNA/RNA sample or PCR product.
Use following amounts of Synthetic carrier RNA:
Recommended final concentration in precipitation solution is 10–20 μg/ml
For example, add 1 μl of 10 mg/ml Synthetic Carrier RNA into 200 μl of RNA
or DNA sample which will be precipitated with 3 x volumes of ethanol.
Alternatively, add 5 μl of 1 mg/ml Synthetic Carrier RNA into 100 μl of RNA
or DNA sample which will be precipitated with 3 x volumes of ethanol.
Incubate at low temperatures: After adding the coprecipitant and precipitation reagents,
incubate the reaction mixture at a low temperature, usually -20°C or lower,
for an adequate duration (e.g., 30 minutes to overnight).
For best purification results, and highest purity PCR purification product, wash the precipitated DNA pellet:
After centrifugation and removal of the supernatant, wash the DNA pellet with ice-cold 70% ethanol
to remove residual impurities and dry the pellet before resolving it in nuclease free water.
Applications
All molecular biology applications where concentration of RNA or DNA solutions is required, such as:
RNA extraction/isolation procedures
DNA extraction/isolation procedures
Clean-up and reprecipitation of RNA or DNA
Benefits
Animal and yeast-free aqueous solution of synthetic RNA
Inert coprecipitating agent helping to increase the concentration of target nucleic acids in low-concentrated solutions
Shows no inhibition in RT-PCR, PCR and qPCR reactions
Stable - can be stored and shipped at ambient temperature