DNA Lysis Buffer A powerful combination of organic and inorganic salts with a unique set of detergents
Effective lysis of prokaryotic and eukaryotic cells
Powerful protein denaturant
Neutralizes polyphenols and polysaccharides
Binding nucleic acids to silica membranes and silica coated magnetic beads
Catalog Number Quantity Price
DLB-01-50 50 ml $ 22.00
DLB-01-100 100 ml $ 44.00
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DNA Bind Buffer A unique combination of organic salts with a proprietary solvent
Proprietary mix of salts and solvents
Binds nucleic acids to silica membranes and silica-coated magnetic beads
Add two volumes of Bind Buffer to one volume of nucleic acid solution
Selective binding of nucleic acids in the presence of polyphenols and polysaccharides
Catalog Number Quantity Price
DBB-01-50 50 ml $ 15.00
DBB-01-100 100 ml $ 30.00
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Wash Buffer I A Wash Buffer to clean-up your phenolic contaminants
Proprietary mix of salts and solvents to clean-up phenolic contaminants
Retains nucleic acids on silica membranes and silica-coated magnetic beads
Washes away phenolic compounds and humic acid related compounds
Effective in washing nucleic acid pellet precipitated in ethanol precipitation
Catalog Number Quantity Price
WB-01-50 50 ml $ 18.00
WB-01-100 100 ml $ 32.00
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Protocols
Whole Blood - Direct Lysis
1. Add 50 to 300 µl of blood to a 2 ml collection tube.
2. Add 800 µl of Lysis Buffer (Cat. # DLB-01) and invert the tubes several times to mix. Vortex for a few seconds to ensure a homogeneous mixture.
3. Add 1 ml of Bind Buffer (Cat. # DBB-01) and invert the tubes at least 5 times to ensure an uniform mixture.
4. Load up to 650 µl on to spin columns and centrifuge at 10,000 x g for 1 minute and discard the flow through.
5. Add remainder of lysate and centrifuge at 10,000 x g for 1 minute. Discard the flow through, retain spin columns and move it back to its collection tube.
6. Wash spin columns using 500 µl of Wash Buffer II (70% ethanol). Load 500 µl of Wash Buffer II and centrifuge the tubes at 10,000 x g for 1 minute. Discard flow through.
7. Place empty spin column in the collection tube and centrifuge one more time at 10,000 x g for
1 minute.
8. Transfer spin column to a new 1.5 ml collection tube and add up to 50 or 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA).
9. Centrifuge the tubes at 10,000 x g for 1 minute.
10. Discard spin columns and store eluted DNA as per your requirements.
Whole Blood - Buffy Coat
1. Add 50 to 300 µl of blood to a 2 ml collection tube followed by the addition of 1.5 ml of
RBC Lysis Buffer.
2. Invert the tubes several times to mix and incubate at room temperature for 5 minutes. Centrifuge at 10,000 x g for 1 minute and discard the supernatant.
3. Resuspend the pellet in 500 µl of Lysis Buffer (Cat. # DLB-01). Pipette up and down to resuspend the pellet completely. Add 500 µl of Bind Buffer (Cat. # DBB-01) and mix by inverting the tubes at least
5 times.
4. Load up to 650 µl onto a spin column and centrifuge at 10,000 x g for 1 minute and discard the flow
through.
5. Add remainder of the lysate and centrifuge again at 10,000 x g for 1 minute. Discard the flow through,
retain the spin column and move it back to its collection tube.
6. Wash the spin columns using 500 µl of Wash Buffer II (70% ethanol). Load 500 µl of Wash Buffer II and
centrifuge the tubes at 10,000 x g for 1 minute. Discard the flow through.
7. Place empty spin column in the collection tube and centrifuge one more time at 10,000 x g for
1 minute.
8. Transfer spin column to a new 1.5 ml collection tube and add up to 50 or 100 µl of Elution Buffer
(Sterile water or 10 mM Tris-5 mM EDTA).
9. Centrifuge the tubes at 10,000 x g for 1 minute.
10. Discard spin column and store eluted DNA as per your requirements.
Body Fluids - Sputum, Unrine etc.
1. Add up to 550 µl of Lysis Buffer (Cat. # DLB-01) in to a 1.5 ml snap cap tube and follow this with the
addition of up to 150 µl of sample.
2. Vortex for a few seconds and incubate at room temperature for 5 minutes.
3. Add 550 µl of 100% ethyl alcohol or Bind Buffer (Cat. # DBB-01) and invert the tubes at least 5 times.
4. Load 650 µl of the solution from step # 3 to spin columns, centrifuge at 10,000 x g for 1 minute and
discard the flow through.
5. Load 650 µl of the solution from step # 3 to spin columns, centrifuge at 10,000 x g for 1 minute and
discard the flow through.
6. Centrifuge the empty spin columns at 10,000 x g for 30 seconds.
7. Wash spin columns twice with 500 µl of 70 % ethanol or DNA Bind Buffer (Cat. # DBB-01). Add
500 µl of 70 % ethanol to spin columns and centrifuge at 10,000 x g for 1 minute. Discard the filtrate and repeat this step one more time.
8. Centrifuge empty spin columns at 10,000 x g for 1 minute and transfer them to a new collection tube.
9. Add 50 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA).and incubate at room
temperature for 1 minute.
10. Centrifuge at 1000 x g for 1 minute and discard spin columns. DNA is ready for all your down stream applications.
Stool Samples
1. Add 10 - 20 mg of fecal sample in to Bead Tubes and add 800 µl of Lysis Buffer (Cat. # DLB-01). Vortex the tubes for about 10 seconds to resuspend samples.
2. Homogenize fecal samples either by vortexing for 5 minutes or using a bead beater as per
manufacturers’ instructions.
3. Centrifuge Bead Tubes at 10,000 x g for 1 minute and transfer the supernatant to a pre filtration column (Cat. # MCP-01). Centrifuge one more time at 10,000 x g for 5 minutes and collect the
flow-through.
4. Discard the pre filter column and the pellet while trasnfering the flow through to a 2 ml collection
tube.
5. Add 1000 µl of Bind Buffer (Cat. # DBB-01) and mix by inverting at least 10 times.
6. Load up to 650 µl to a spin column and centrifuge at 10,000 x g for 1 minute. Discard the flow
through and, process the remaining volume and discard the flow through again.
7. Wash the spin columns twice, with Wash buffers I & II, respectively. Add 500 µl of Wash buffer I
(Cat. # WB-01) and centrifuge for 1 minute at 10,000 x g and discard flow through. Add 500 µl of
Wash Buffer II (70% ethanol) and centrifuge at 10,000 x g for 1 minute. Discard the flow through.
8. Centrifuge empty spin columns at 10,000 x g for 1 minute and remove the residual Wash Buffers and
transfer the spin column (discard the collection tube) to a 1.5 ml collection tube.
9. Add 50 - 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge one more
time at 10,000 x g for 1 minute. Discard the spin column and store eluted DNA as per your
requirements.
Mammalian Tissues
1. Add 10 - 20 mg of tissue samples in to Bead Tubes and add 800 µl of Lysis Buffer (Cat. # DLB-01). Vortex
the tubes for about 10 seconds to resuspend samples. Depending on tissue types, you may have
to use a mechanical homogenizer.
2. Homogenize samples either by vortexing for 5 minutes or using a bead beater as per manufacturers’
instructions.
3. Centrifuge Bead Tubes at 10,000 x g for 1 minute and transfer the supernatant to a 1.5 ml collection
tube. Centrifuge one more time at 10,000 x g for 5 minutes and transfer the supernatant to a 2 ml
collection tube.
4. Add 1000 µl of Bind Buffer (Cat. # DBB-01) and mix by inverting at least 10 times.
5. Load up to 650 µl to a spin column and centrifuge at 10,000 x g for 1 minute. Discard the flow
through and process the remaining volume and discard the flow through again.
6. Wash the spin columns with Wash buffer II (70% ethanol). Add 500 µl of Wash buffer II and
centrifuge for 1 minute at 10,000 x g and discard flow through.
7. Centrifuge empty spin columns at 10,000 x g for 1 minute and remove the residual Wash Buffers and
transfer the spin column (discard the collection tube) to a 1.5 ml collection tube.
8. Add 50 - 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge one more
time at 10,000 x g for 1 minute. Discard the spin column and store eluted DNA as per your
requirements.
Microorganisms
1. Centrifuge 1 ml of an overnight culture at 10,000 x g for 1 minute. Discard the supernatant and
centrifuge again at 10,000 x g for 1 minute. Remove the residual culture liquid, leaving the pellet
intact.
2. Add 800 µl of Lysis Buffer (Cat. # DLB-01) and resuspend the microbial pellet. Transfer the
suspension to a bead tube and homogenize the tubes either by vortexing for 5 minutes or using a
bead beater as per manufacturers’ instructions.
3. Centrifuge Bead Tubes at 10,000 x g for 1 minute and transfer the supernatant to a 1.5 ml collection
tube. Centrifuge one more time at 10,000 x g for 5 minutes and transfer the supernatant to a 2 ml
collection tube.
4. Add 1000 µl of Bind Buffer (Cat. # DBB-01) and mix by inverting at least 10 times.
5. Load up to 650 µl to a spin column and centrifuge at 10,000 x g for 1 minute. Discard the flow
through and process the remaining volume and discard the flow through again.
6. Wash the spin columns with Wash buffer II (70% ethanol). Add 500 µl of Wash buffer II and
centrifuge for 1 minute at 10,000 x g and discard flow through.
7. Centrifuge empty spin columns at 10,000 x g for 1 minute and remove the residual Wash Buffers and transfer the spin column (discard the collection tube) to a 1.5 ml collection tube.
8. Add 50 - 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge one more
time at 10,000 x g for 1 minute. Discard the spin column and store eluted DNA as per your
requirements.
Plants
1. Add up to 50 mg of tissue samples in to Bead Tubes and add 800 µl of Lysis Buffer (Cat. # DLB-01).
Vortex the tubes for about 10 seconds to resuspend samples. Depending on the tissue types, you
may have to use a mechanical homogenizer.
2. Homogenize tissue samples either by vortexing for 5 minutes or using a bead beater as per
manufacturers’ instructions.
3. Centrifuge Bead Tubes at 10,000 x g for 1 minute and transfer supernatant to a 1.5 ml collection tube.
Centrifuge one more time at 10,000 x g for 5 minutes and transfer the supernatant to a 2 ml collection
tube.
4. Add 1000 µl of Bind Buffer (Cat # DBB-01) and mix by inverting at least 10 times.
5. Load up to 650 µl to a spin column and centrifuge at 10,000 x g for 1 minute. Discard the flow
through and process the remaining volume and discard the flow through again.
6. Wash the spin columns with Wash buffer II (70% ethanol). Add 500 µl of Wash buffer II and
centrifuge for 1 minute at 10,000 x g and discard flow through.
7. Centrifuge empty spin columns at 10,000 x g for 1 minute and remove the residual Wash Buffers and
transfer the spin column (discard the collection tube) to a 1.5 ml collection tube.
8. Add 50 - 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge one more
time at 10,000 x g for 1 minute. Discard the spin column and store eluted DNA as per
your requirements.
Seeds
1. Add up to 20 mg of ground seed samples in to Bead Tubes and add 800 µl of Lysis Buffer
(Cat. # DLB-01). Vortex the tubes for about 10 seconds to resuspend samples. Depending on the
seed type, you may have to use a mechanical homogenizer.
2. Scale down the starting material depending on seed composition.
3. Centrifuge Bead Tubes at 10,000 x g for 1 minute and transfer the supernatant to a 1.5 ml collection
tube. Centrifuge one more time at 10,000 x g for 5 minutes and transfer the supernatant to a 2 ml
collection tube.
4. Add 1000 µl of Bind Buffer (Cat # DBB-01) and mix by inverting at least 10 times.
5. Load up to 650 µl to a spin column and centrifuge at 10,000 x g for 1 minute. Discard the flow
through and process the remaining volume and discard the flow through again.
6. Wash the spin columns with Wash buffer II (70% ethanol). Add 500 µl of Wash buffer II and
centrifuge for 1 minute at 10,000 x g and discard flow through.
7. Centrifuge empty spin columns at 10,000 x g for 1 minute and remove the residual Wash Buffers and
transfer the spin column (discard the collection tube) to a 1.5 ml collection tube.
8. Add 50 - 100 µl of Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge one more
time at 10,000 x g for 1 minute. Discard the spin column and store eluted DNA as per your
requirements.
Soil Samples
1. Add up to 500 mg of soil samples in to Bead Tubes and add 1200 µl of Lysis Buffer (Cat. # DLB-01).
Vortex for about 10 seconds to resuspend the soil mass.
2. Homogenize soil samples either by vortexing for 5 minutes or using a bead beater as per
manufacturers’ instructions.
3. Centrifuge Bead Tubes at 7,000 x g for 1 minute and transfer the supernatant to a 1.7 ml collection
tube.
4. Centrifuge at 10,000 x g for 5 minutes and transfer the supernatant to a 2 ml collection tube.
5. Add 1000 µl of Bind Buffer (Cat. # DBB-01) and mix by inverting at least 10 times.
6. Load lysate directly on to spin columns and process the entire volume by centrifuging at
10,000 x g for 1 minute.
7. Wash the columns with 500µl of Wash Buffer I (Cat. # WB-01) and discard flow through. Repeat this
step with Wash Buffer II (70% ethanol).
8. Centrifuge the empty spin baskets at 10,000 x g for 1 minute to remove salts.
9. Transfer the empty spin baskets to a new 1.7 ml collection tube and add 50 to 100 µl of
Elution Buffer (Sterile water or 10 mM Tris-5 mM EDTA) and centrifuge at 10,000 x g for 1 minute.
Discard the spin basket and store DNA as per your requirements.