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DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) are essential biomolecules that act as carriers of genetic information and play a central role in the biology of all living organisms. DNA stores the genetic information necessary for the development, function and reproduction of organisms, while RNA serves as a mediator in the transmission of this information and in protein synthesis. The structural differences between DNA and RNA lie in their sugar components and the bases they contain, which gives them different functions in cell metabolism.
High performance liquid chromatography (HPLC) is an analytical method commonly used to separate, identify and quantify DNA and RNA components. This technique is particularly important as it offers high precision and sensitivity, which is crucial for analysing nucleic acids in research and diagnostics. By using HPLC, researchers can efficiently analyse specific nucleotides, nucleosides and their modifications, which is essential for understanding genetic processes and developing therapeutic approaches.
The terms nucleotide, oligonucleotide and DNA/RNA refer to different molecular sizes in the analysis of nucleic acids that can be analysed by HPLC.
1. 100nt 2. 200nt 3. 300nt 4. 400nt 5. 500nt
Column: RiboSep RNA 50x7.8mm, 2µm
Mobile phase: A: 0.1M TEA; B: 0.1M TEAA, 25% ACN
Isocratic: 15min
Injection volume: 10µL
1. n-2 ( 5'-AAA AGT CCG TGA GA-3' ; 16.4 min)
2. n-1 ( 5'-CAA AAG TCC GTG AGA-3' ; 17.1 min)
3. 16mer (5'-ACA AAA GTC CGT GAG A-3' ; 17.8 min)
Test conditions:
A: 0.025 M tetrabutylammonium bromide (TBuABr)
B: 0.025 M TBuABr, 50% CH3CN; 10-90% B in 17 min, 60-100% B in 1 min, 100-10% B in 1 min
Temperature: 60°C
Injection volume: 5 µL
UV detection: 260 nm
siRNA - Reversed Phase:
DNA - Reversed Phase:
DNA - SEC:
DNA fragments - IEX:
RNA - IEX:
tRNA, rRNA, DNA - IEX:
DNA - IEX:
DNA - SEC:
DNA - Reversed Phase:
DNA Digest - IEX:
CNT-DNA - HILIC:
dsRNA, mRNA, ssRNA - Reversed Phase:
dsRNA, ssRNA - SEC:
DNA - Reversed Phase:
DNA - IEX:
DNA - SEC:
RNA - IEX:
RNA - SEC:
mRNA - Reversed Phase:
mRNA - SEC:
mRNA - affinity chromatography:
circular RNA - Reversed Phase:
circular RNA - SEC:
circular mRNA - SEC:
mRNA - Reversed Phase:
tRNA - IEX:
mRNA - Reversed Phase:
mRNA - IEX:
DNA - Reversed Phase:
DNA - IEX:
dsDNA - IEX:
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