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Example sentences for: novagen
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To assemble Net1N beads with Cdc14 for six reactions (Figure 4A, detection method: Coomassie Blue staining), 105 μl of anti-T7 beads (Novagen) were incubated with 24 μg of His6-T7-Net1N in 100 μl of HEBD for 1 hr, washed four times with HBS + 0.2% n-octylglucoside + 2 mM DTT, and incubated with 6 μg of GST-Cdc14 in 100 μl of HEBD for 1 hr.
Protein induction was performed using isopropyl-β-D-thiogalactopyranoside (IPTG) as described by Novagen Inc.
The wild type autogene thus obtained (pET/T7p/T7) was transformed into strain HMS174 pLysS (Novagen, Madison, WI), the same cell line Studier et al.
We used the Quickchange kit (Stratagene) to generate site directed mutants, V80R and A61D from wild-type TEL-SAM cloned into a modified pET3c vector (Novagen) [ 7 ] . The expressed protein sequence includes an MEKTR leader sequence, followed by residues 38-124 of the TEL protein and then a C-terminal His tag.
Wax-embedded embryonic sections were purchased from Novagen (Madison, Wl).