DP108-EndoFree Midi Plasmid Kit

EndoFree Midi Plasmid Kit
For purification of ultrapure plasmid DNA with high yield

EndoFree Midi Plasmid Kit
(Spin Column)
Cat. no. 4992853

Storage Condition
EndoFree Midi Plasmid Kit can be stored dry at room temperature (15-30°C)
for up to 15 months. If any precipitate forms in the buffers, it should be
dissolved by warming the buffers at 37°C for several minutes before use.
RNase A can be stored for 15 months at room temperature (15-30°C). After
the addition of RNase A, Buffer P1 should be stored at 2-8°C and it would
be stable for 6 months.

Introduction
EndoFree Midi Plasmid Kit uses unique silica membrane technology which
can specifically adsorb plasmid DNA efficiently. Meanwhile, this kit also
uses unique Buffer EBT and Filtration CS1 to get rid of contaminants like
endotoxin and protein compounds effectively. The whole experimental
procedure of plasmid DNA extraction could be finished within 1 h. Plasmid
DNA prepared by EndoFree Midi Plasmid Kit is suitable for a variety of
downstream applications including restriction enzyme digestion, PCR,
sequencing, ligation, transformation and cell transfection.
Cell culture volume: For the isolation of high copy plasmid DNA, 50 ml of
E. colicell culture is recommended, and the yield would be within 250-750
μg. For the isolation of low copy plasmid DNA, 100 ml of E. colicell culture
is recommended, and the yield would be within 100-300 μg

Important Notes Before Starting

1. Add the provided RNase A solution to Buffer P1 (use 1 vial RNase A per
   bottle Buffer P1), mix, and store at 2-8°C.
2. Add ethanol (96-100%) to Buffer PWF and Buffer MRDE before use,
   check bottle tag for volume.
3. Check Buffer BL, P2, E3 and EBT before use to see if there is any
   precipitate formed. If necessary, dissolve the precipitate by warming at
   37°C for several minutes.
4. Avoid direct contact of Buffer P2, E3 and EBT, immediately close the lid
   after use.
5. Draw out the plunger from the Filtration CS1 slowly to avoid membrane
   loose.
6. The plasmid yield is related to cell concentration and copy number of
   plasmid. If working with low copy vectors or large plasmid (>10 kb), it
   may be beneficial to increase culture volume and to increase Buffer P1,
   P2, E3 and EBT in proportion. Warm the Buffer TB at 65-70°C before
   use (Prolong adsorption and elution time properly could increase
   extraction efficiency).
7. Use Buffer BL to equilibrate spin columns before use could maximally
   activate silica membrane and increase the yield.
8. After treatment with Buffer BL, use the spin column as soon as possible,
   or else the efficiency would be reduced.

DP108-EndoFree Midi Plasmid Kit

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