precipitate in NEB T4 Ligase buffer?

Peter Rudolph p_rudolph at MORPH.SPACENET.DE
Tue Feb 14 19:02:41 EST 1995


 *  In article <3gr9rm$jon at mcmail.CIS.McMaster.CA>
 *  g9013118 at mcmail.cis.mcmaster.ca (K. Legate) writes:
 *
 * |: It's the DTT. However, repeated freeze-thawing of ATP does have effects,
 * |: at least according to one supplier of T4 ligase.
 * |
 * | The efficiency of my ligations goes way down after repeated freeze-thaw
 * | cycles of the buffer. Adding a small amount of ATP to the mix restores
 * | original efficiency.
 * |
 * | kyle
 *
 * ...and how is the rATP stored? I keep mine at -20 C. If yours is also frozen
 * at -20 C, then thawed before adding a small amount to the ligation buffer,
 * wouldn't the rATP also degrade over several freeze-thaw cycles resulting in
 * loss of ligation efficiency?

...in our lab we store the ligation buffers in small aliquots so there's no
need to freeze and thaw the mix several times. You simply thaw the aliquot
an throw away the rest of it...only a idea but it works fine...

PR.





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