AquaPlasmidTM is
a novel
aqueous solution-based reagent for isolation and purification of
plasmid DNA. It yields ~5-10 ug of plasmid DNA from a milliliter of
overnight bacterial culture. The isolated DNA is ultra pure and has an
A260/A280 ratio of 1.8-2.0 and A260/A230 ratio of 2.0-2.2. The
extraction protocol is as simple as alkaline lysis. It may be carried
out using an inexpensive personal picofuge, allowing users to perform
plasmid DNA extraction in resource-limited settings such as high school
classrooms or without needing to fight for a shared
high-speed centrifuge in a busy research lab.
AquaPlasmidrepresents a breakthrough in
plasmid DNA isolation. It allows the isolation and
purification of plasmid DNA without using solid support, such as columns, resins,
slurries, beads, filters, membranes, and cartridges. It greatly
reduces material and energy consumption in producing these products and
the generation of plastic wastes in using these products.
Simple
The AquaPlasmid protocol is as simple as
alkaline lysis: cells are lysed, debris is removed, and plasmid DNA is then precipitated. No
buffer 1, 2, 3, 4, ..., or A, B, C, D, .... Furthermore, the
entire extraction can be done without needing a high-speed centrifuge,
just a personal picofuge will do, enabling plasmid DNA extraction in
high school science classrooms or resource-limited laboratories; and
you could even do 96-well HTP minipreps with a speed-vac centrifuge.
Nontoxic
No phenol, no chloroform, no saturated butanol, no
ethidium bromide, no cesium chloride, no guanidine hydrochloride, or any other toxic component.
Pure
The
isolated plasmid DNA is essentially free of
cellular impurities and enzyme inhibitors. A260/A280 = 1.8-2.0 and
A260/A230 = 2.0-2.2. Excellent for all downstream applications.
Scalable
One
single AquaPlasmid Kit does all: mini, midi,
maxi, and 96-well HTP; no need to purchase separate and bulky mini,
midi, maxi,
and HTP kits.
High Yield
10-20, 100-200, 1000-2000 ug of DNA can be isolated
from 2 ml (mini), 20 ml (midi), 200 ml
(maxi) overnight TB cultures.
Cost Saving
A 15-ml AquaPlasmid kit costs $199, good for 300
minipreps, that is only $0.66/miniprep. It is the most inexpensive plasmid DNA isolation kit on the market.
Comparison
Below is a comparison of AquaPlasmid and column-based
competitors. You can easily see the advantages and benefits of using
AquaPlasmid.
Competitors
AquaPlasmid
Isolation
Mechanism
Selective
Binding
Selective
Extraction
Isolation
Format
Solid
Support
Aqueous
Solution
Price
($/miniprep)
1.50
0.66
Guanidine
HCl
Yes
No
Scalable
No
Yes
#
of Solutions in the Kit
7
2
Yield
(ug DNA/ml culture)
2-4 (LB)
Can't
use TB
3-5 (LB)
6-10 (TB)
Purity
(A260/A280)
1.8-2.0
1.8-2.0
Sequencing
(Phred20)
~700
~900
Brief Protocol
1.Harvest the Cells
Centrifuge 2 ml overnight culture at 3,000-14,000 xg at room temperature for 1 min
to pellet the bacteria. Aspirate to remove the culture medium as completely
as possible.
2.
Lyse the Cells
Add 80 ul of deionized water to the cell pellet.
Vortex vigorously to fully resuspend the cells. Add 100 ul of AquaLysis solution to the
cell suspension. Vortex vigorously to mix the contents well. Incubate at room
temperature for 5 min.
3.
Remove the Debris
Add 50 ul of AquaPlasmid solution (15 ml in the kit
for 300 minipreps) to the crude lysate. Touch-vortex (a few seconds on and
then a few seconds off) at top speed or shake vigorously 10-15 times to mix
the contents. Incubate at room temperature for 5 min. Centrifuge at 3,000-14,000 xg for 5 min to pellet the debris.
4.
Pellet the Plasmid DNA
Transfer
the clear lysate (210 ul) to a new 0.5-ml
microfuge tube. Add 1/3 vol (70 ul) of isopropanol to the
lysate. Vortex for 20-30 sec to mix well. Centrifuge at 3,000-14,000 xg for
5 min to pellet the plasmid DNA. Flip the tube to discard the supernatant.
Fill the tube with 70% ethanol, and then flip to discard the ethanol. Repeat
the ethanol rinse once. Air-dry the DNA pellet (nearly invisible) for ~5 min.
Suspend the DNA in 50 ul of deionized water. The
expected DNA yield is 5-10 ug per ml of overnight culture.
The DNA purity is ~1.8-2.0 at 260/280 and ~2.0-2.2 at 260/230.
Ordering Information
Product
Name: AquaPlasmidTM Kit
Product
Number: 1001, 1015
Application:
Isolation of plasmid DNA. For in
vitro research use only.
Size:
The kit is sufficient for the preparation of
1001: 10
minipreps (use 50 ul AquaPlasmid for 2 ml overnight culture)
1015: 300 minipreps, or 30 midipreps, or 3 maxipreps
Kit
Contents: The AquaPlasmid Kit includes the
following items
1001: 0.5 ml AquaPlasmid solution, 1 ml AquaLysis solution, User Manual
1015: 15 ml AquaPlasmid solution, 30 ml AquaLysis solution, User Manual
Price:
1001: $10 each
1015: $199 each
Ordering:
To order, please
click the "BUY" button below.
