effect of TE buffer on cells with cell lysis solution - (Dec/12/2006 )
hello. i am going to isolate dna from buccal swabs. I however accidentally placed TE buffer on the microcentrifuge tube with the collection brush because I have not obtained the kit yet.
The kit said that i need to place Cell Lysis Solution to the tube with collection brush instead of the TE buffer.
Will there be an effect if I combine the two solutions and just go on and still follow the protocol of the kit? ( i still place cell lysis solution even if there is a TE buffer already?)
or should i spin down the cells together with the TE component and remove the collection brush and discard the TE supernatant and add the cell lysis buffer and continue with the described kit protocol?
It might work with just combining them, but the safer version is to spin down and resuspend in the correct buffer. A possible issue is the bursting of cells in TE which has low osmotic strength. If the cells are not intact, then spinning them down will not recover the DNA you need. A third possibility is to add more than 50% lysis buffer, or to add lysis components directly. SDS added to the TE would certainly lyse cells, but may affect the downstream uses.
thank you so much. if i placed my sample in cold temperature, will it minimize the osmotic pressure and bursting of cells and in that case i can minimize the lost of my dna? i plan to spin down and remove the TE and replace with cell lysis. thanks once again.