Low melt temp agarose not melting - (Jan/24/2006 )
hi everyone...once more again i have cut the piece of my digested vector and am trying to disolve in it buffer.....as molecular cloning technique says.....buit its not melting at 65....can I try higher temperatures? I have used low melting temperature agarose but ....
what would you do?
Buy a new bottle of low melting temperature agarose and try again.
does it mean the gel is not good...??? 
but i have already purified the fragment please is there any way to melt it? i can pipette it up and down somehow but corners of agarose piece are seen also... 
does it mean that it has melted or not melted????
what percentage agarose did you use to make the gel?
1%.....too much?
no 1% should be fine. I am not sure why its not melting. Are you leaving it long enough? I presume you are shaking the tube form time to time? It shouldn't be hard to dissolve though.
what percentage agarose did you use to make the gel?
1%.....too much?
This is from the earlier post where you can't use the freeze n squeeze method right?? you are dissolving in what buffer exactly?? sorry will try to find other post....
im disolving in LMT buffer (Tris and EDTA) and at 65 for 5 minutes.....no this is the first time im posting this problem....no other post.....i dont know im able to pipet it....it seems very loose but still I can see some agarose still not melted....what is the squeeze and freeze method? can I use it in my case?
no im not shaking...
....and actually my tube is little bit bigger than the hole of the heating block...so the lid is not closing comletly....maybe that is the problem... ...how do u heat the 1.5 ml ependorf to 65? 
okay that one wasn't you... I like the freeze n squeeze no binding and you can get the ones that remove EtBr too... just freeze the gel piece smash it up and centrifuge into a 1.5mL tube then etoh ppt if necessary... DNA and TAE from gel go through, the agarose stays on top of the filter, EtBr is filtered out if you get the minus etbr columns... no melting either...