So I have been running native PAGE (15%) for my DNA complexes, from what I observe, there is only one DNA complex as shown in image, 'RQ' which forms partially ds DNA always has some streaking issue. Could anyone suggest what it could be...is it impurtities or unwanted complexes forming.
Strand length of R is a single strand with 49 bases, Q also is a single strand with 32 bases. Oligos are HPSF purified.
Your R samples suggest that the wells are well rinsed and clean so it is probably not a loading issue. You could get this effect if the RQ can form 2 complexes which are in equilibrium and melting and reforming at the gel running temperature so a snapshot view of the gel appears to show the dna running at many different speeds so looks smeared. It may be that running the gel at a different temperature could favour one of the heteroduplex structures and could change the pattern that you see
Thank you for your response Paul Rutland . Currently, I am running the gel at 4 °C. So then I should try it at room temperature? Is that your suggestion?
I do not know what temperature is best for stabilising the wanted form but I think that the temperature of a PAGE gel is determined largely by the applied voltage and the amounts of salts in the buffer so I would try running at a higher voltage first and if this does not improve the gel then at a lower voltage than your current running conditions Keziah Souza
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Inorganic Materials
- Ceramics
- Glass