Figure 4. High salt buffers arrest translation after cell lysis better than Cm.
(A) We added [14C]Lys-tRNALys to frozen lysate that was then thawed for 15 min. [14C]Lys that was incorporated into nascent peptides can be selectively precipitated with TCA after tRNA hydrolysis under alkaline conditions. (B) Lysates were made with buffers containing 1 mM Cm, 50 mM MgCl2, or 1M NaCl. The boiled sample was denatured prior to addition of Lys-tRNA. Error bars reflect the standard deviation of four technical replicates. The boiled and Cm samples were compared using a one-tailed, paired t-test.