Invitrogen Bac to Bac Expression System
Invitrogen Bac to Bac Expression System
Invitrogen Bac to Bac Expression System
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Isolating Recombinant <strong>Bac</strong>mid DNA, continued<br />
DNA Isolation<br />
Procedure,<br />
continued<br />
12. Remove as much of the supernatant as possible, taking care not <strong>to</strong> disturb the<br />
pellet. Air dry the pellet for 5 <strong>to</strong> 10 minutes at room temperature. Do not<br />
overdry the pellet. Important Note: Do not use a SpeedVac <strong>to</strong> dry the DNA<br />
pellet as this may shear the DNA.<br />
13. Dissolve the DNA pellet in 40 µl of 1X TE Buffer, pH 8.0. To avoid shearing,<br />
do not mechanically resuspend the DNA. Allow the solution <strong>to</strong> sit in the tube<br />
with occasional gentle tapping of the bot<strong>to</strong>m of the tube.<br />
14. S<strong>to</strong>re the DNA at +4°C. Note: We do not recommend s<strong>to</strong>ring the purified<br />
bacmid DNA at -20°C as repeated freezing and thawing may shear the DNA.<br />
15. Proceed <strong>to</strong> analyze the recombinant bacmid DNA (see Analyzing<br />
Recombinant <strong>Bac</strong>mid DNA by PCR, page 23) or <strong>to</strong> transfect the DNA in<strong>to</strong><br />
insect cells (see Transfecting Insect Cells, page 26).<br />
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