DNA - RNA

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.

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Technical Data

Differences between nucleotides, oligonucleotides and DNA/RNA

The terms nucleotide, oligonucleotide and DNA/RNA refer to different molecular sizes in the analysis of nucleic acids that can be analysed by HPLC.

 

  • Nucleotides are the basic building blocks of DNA and RNA, consisting of a base, a sugar and a phosphate group. HPLC analysis of nucleotides involves identifying and quantifying individual nucleotides. This analysis can be used, for example, to determine the content of specific nucleotides in a sample or to analyse their modifications.
  • Oligonucleotides are short chains of nucleotides, typically up to about 20-30 base pairs in length. HPLC is used to check the purity, composition and sequence of these short DNA or RNA fragments. This analysis is particularly important in the production of synthetic oligonucleotides used in research or medicine.
  • DNA/RNA analysis using HPLC involves analysing longer DNA or RNA strands, which can comprise several hundred to thousands of base pairs. In this analysis, structure, modifications and fragmentation patterns can be analysed. For long strands, HPLC is often used in combination with other techniques such as mass spectrometry or gel electrophoresis to analyse the complete sequence or structure.

Applications

Separation of different RNA chain lengths with Concise RiboSep RNA 50x7.8mm, 2µm

Peak identities

1. 100nt 2. 200nt 3. 300nt 4. 400nt 5. 500nt

Test conditions

Column: RiboSep RNA 50x7.8mm, 2µm

Mobile phase: A: 0.1M TEA; B: 0.1M TEAA, 25% ACN

Isocratic: 15min

Injection volume: 10µL

Separation of different DNA oligonucleotides with ADS Biotec DNASep

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)

Downloads







Sepax

DNA - Reversed Phase:

  • Proteomix RP-300, 10µm

 

DNA - IEX:

  • Proteomix WAX, 5µm

 

DNA - SEC:

  • Zenix SEC-300, 3µm

 

RNA - IEX:

  • Proteomix WAX, 5µm

 

RNA - SEC:

  • SRT SEC-2000, 5µm

 

mRNA - Reversed Phase:

  • Proteomix RP-1000, 5µm

 

mRNA - SEC:

  • Zenix SEC-300, 3µm
  • SRT SEC-1000, 5µm

 

mRNA - affinity chromatography:

  • Monomix dT(20), 30µm

 

circular RNA - Reversed Phase:

  • Bio-C18, 5µm

 

circular RNA - SEC:

  • SRT SEC-2000, 5µm

 

circular mRNA - SEC:

  • SRT SEC-1000, 5µm

Shimadzu

mRNA - Reversed Phase:

  • Shim-pack Scepter Claris C18, 1.9µm

Shodex

tRNA - IEX:

  • IEC DEAE-825



Waters

DNA - Reversed Phase:

  • ACQUITY Peptide Separation Technology BEH300 C18, 1.7µm

 

DNA - IEX:

  • Protein-Pak Hi Res Q, 5µm

 

dsDNA - IEX:

  • Protein-Pak Hi Res Q, 5µm

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