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Each sample was spotted into the block twice.
When this strain was spotted into a medium allowing the expression of m8 and Hsp104, the growth was clearly impaired (Figure 5A).
Negative control siRNAs (RISC-free siRNA #4390843 (Ambion) and NegsiRNA #1022076 (Qiagen, Valencia, CA) were also spotted into four independent wells each.
A total of 12.5 ng siRNA in 1 3 µl of siRNA in suspension Buffer/RNase-free water was spotted into a single well of a 96-well plate.
Briefly, 2 µl of each siRNA at a concentration of 1 µM was spotted into four independent wells of a 384-well dish and stored at −80°C until use.
The digested tryptic peptides were then mixed with CHCA matrix (alpha-cyano-4-hydroxycinnamic acid) and spotted into wells of a MALDI plate for MALDI-TOF MS identification (ABI4800, Applied Biosystems, Foster City, CA).
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