which codes the JEss MGKRSAGSIMWLASLAVVIACAGA,
and the underlined is the Kozak sequence for efficient expression). Then the ligation is transformed into TOP10 competent
cells and correct clones confirmed by restriction enzyme digestion after mini-prep (see Note 3). A glycerol stock is then
generated for future use.
3. Every time when beginning the Mega or Giga prep, always start
with a single colony from either a new transformation or by
restreaking of glycerol stock. Pick a single colony and inoculate
in 10 mL 2YT medium and shake at 220 rpm, 37
C for
12–16 h.
Fig. 2 Plasmid map of pcDNA3-JEss DNA vaccine vector
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Lei Li et al.
and the underlined is the Kozak sequence for efficient expression). Then the ligation is transformed into TOP10 competent
cells and correct clones confirmed by restriction enzyme digestion after mini-prep (see Note 3). A glycerol stock is then
generated for future use.
3. Every time when beginning the Mega or Giga prep, always start
with a single colony from either a new transformation or by
restreaking of glycerol stock. Pick a single colony and inoculate
in 10 mL 2YT medium and shake at 220 rpm, 37
C for
12–16 h.
Fig. 2 Plasmid map of pcDNA3-JEss DNA vaccine vector
412
Lei Li et al.