Biology Forum › Genetics › DNA backbone breakage
- AuthorPosts
- October 4, 2010 at 4:39 pm #13873
NewyorkerParticipantHi,
As DNA denatures as a result of being heated up, could the backbone also break, or is it just the two strands that separate?
Are there any instances where the DNA backbone does break? I know that the bonds between the phosphates and OHs are very strong, and you always hear about the bases separating but not the backbone (which is on the outside, so it should make sense that the separating force will get to it first). When IS the DNA backbone attacked? Have there been studies done that specifically target the strength of the DNA backbone and its ability to hold out against damaging forces?
Thank you! 🙂
-Newyorker
- October 4, 2010 at 5:37 pm #101631canalonParticipant
Just heating should normally not degrade the backbone, so only the 2 strands will separate. In order to cut the backbone:
– restriction enzymes will cut it in specific sites
– shearing forces or other mechanical forces can do it too (at no specific site though), that is why it is not recommended to vortex minipreps after the lysis step. - October 5, 2010 at 3:54 am #101640kanagasundarParticipant
DNA consist of three component i.e., Sugar (deoxyribose), Base ( Purine and Pyridine), Phosphate group. DNA backbone made up of alternative sugar and phosphate are held together by Phosphodiester bond. Phosphodiester bond is a covalend bond, so energy is required to more for breaking phosphodiester bond. Two strands are held together by weak bond i.e., "Hydrogen bond". This bond can easily disturb by heat, as compare to DNA backbone. In the phosphodiester bond has phosphate group bear negative charge. This -ve charge compensated by basic protein histone (made up of arginine And lysine and histine.) When their "ph" change disturb the DNA backbone. But i don’t know that fully disturb, I think not fully disturb because presence of phosphodiester bond.
Various restriction enzyme cut DNA backbone at specific sites.
- October 5, 2010 at 11:13 am #101647JackBeanParticipant
if you’re talking about the standard denaturation in PCR like 95°C for 3 minutes, that should be fine, of course, prolonged heating or even higher temperatures could cause some damage
- October 5, 2010 at 1:29 pm #101648kanagasundarParticipant
Is any method there for separating DNA backbone without damage or little damage?
- October 5, 2010 at 1:51 pm #101651canalonParticipantquote kanagasundar:Is any method there for separating DNA backbone without damage or little damage?
separate the back bone from what?
OPening the double helix? just heat
Removing the bases from the backbone? No clue…
- AuthorPosts
You must be logged in to reply to this topic.
