RNA 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, making protein precipitation unnecessary
Inactivates nucleases
Neutralizes polyphenols and polysaccharides
Binding nucleic acids to silica membranes and silica coated magnetic beads
Catalog Number Quantity Price
RLB-01-50 50 ml $ 25.00
RLB-01-100 100 ml $ 45.00
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RNA 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 one volume of Bind Buffer to one volume of RNA solution
Selective binding of nucleic acids in the presence of polyphenols and polysaccharides
Use instead of 100% ethanol for binding to silica membranes and silica-coated magnetic beads
Catalog Number Quantity Price
RBB-01-50 50 ml $ 17.00
RBB-01-100 100 ml $ 34.00
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Protocols
Whole Blood - Buffy Coat
1. Add up to 2 ml of blood in a 15 ml collection tube and add 10 ml of 1x RBC Lysis Buffer. Invert the
tubes several time and incubate on ice for 10 minutes, or at 4°C for 10 minutes.
2. Centrifuge at 5000 rpm for 10 minutes and discard the supernatant. Rinse the pellet with
1 ml of 1x RBC Lysis Buffer by adding slowly and inverting the tube once and discarding
1x RBC Lysis Buffer.
3. Resuspend the pellets in 1.8 ml of 1x RBC Lysis Buffer and incubate on ice or at 4°C for 10 minutes. Centrifuge at 10,000 x g for 1 minute and discard the supernatant.
4. Resuspend the pellet in 1 ml of 1x RBC Buffer, add another 800 µl of 1x RBA Buffer and invert gently
5 times.
5. Centrifuge at 10,000 x g for 1 minute and discard the supernatant.
6. Resuspend the pellet in 900 µl of RNA Lysis Buffer (Cat. # RLB-01).
7. Add 900 µl of RNA Bind Buffer (Cat. # RBB-01) and invert the tubes several times to get a
homogeneous mixture.
8 . Load 650 µl onto a spin column and centrifuge at 10,000 x g for 1 minute. Process the entire volume,
it should take a total of three spins.
9. Wash the spin columns with 500 µl of Wash Buffer. Add 500 µl of Wash Buffer and centrifuge at
10,000 x g for 1 minute. Discard the flow through and centrifuge the empty basket one more time at 10,000 x g.
10. Transfer the empty basket to a new 1.7 ml collection tube and add 50 µl of Elution Buffer (Sterile water). Centrifuge at 10,000 x g for 1 minute.
11. Discard spin column and store eluted RNA as per your requirements.
Body Fluids
1. Add 50 to 300 µl of body fluid to a 2 ml collection tube followed by the addition of 800 µl of RNA
Lysis Buffer (Cat. # RLB-01).
2. Incubate at room temperature for 5 minutes and invert the tubes to mix at least once during this
incubation. Centrifuge at 10,000 x g for 1 minute and remove the supernatant to a 2 ml tube.
3. Add 800 µl of RNA Bind Buffer (Cat. # RBB-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).
9. Centrifuge the tubes at 10,000 x g for 1 minute.
10. Discard spin column and store eluted RNA as per your requirements.
Stool Samples
1. Add 10 - 20 mg of fecal sample in to Bead Tubes and add 800 µl of Lysis Buffer (Cat. # RLB-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 transfering the flow through to a 2 ml collection
tube.
5. Add 1000 µl of Bind Buffer (Cat. # RBB-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) 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. # RLB-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. # RBB-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) 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. # RLB-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. # RBB-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) 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. # RLB-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 # RBB-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) 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. # RLB-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 # RBB-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) 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. # RLB-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. # RBB-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) and centrifuge at 10,000 x g for 1 minute. Discard the spin basket and
store DNA as per your requirements.