Direct Extraction Buffer
Buffer for fast extraction of DNA and RNA from sample material
For general laboratory use.
Shipping: shipped at ambient temperature
Storage Conditions: store at 4°C or -20°C
Shelf Life: 12 months
Form: liquid
Concentration: 10 x
Description:
Direct Extraction Buffer allows an easy and fast extraction of DNA and RNA directly from blood, swabs and animal- or plant tissue. The buffer is optimized for use in combination with Direct PCR or RT-PCR master mixes like qPCR ProbesMaster (PCR-396) or SCRIPT Direct RT-qPCR ProbesMaster (PCR-528).
The mix allows DNA and RNA preparation within 3-5 minutes and with a minimum of pipetting steps. It is especially recommended for:
- Direct detection of viral or bacterial DNA in nasal or throat swabs
- Direct PCR from blood samples
- Direct amplification of target DNA from various tissue samples
- Point-of-Care diagnostics
The preparation process can be easily automatized.
Content:
Direct Extraction Buffer
10 x conc.
Sample preparation
a) Blood Samples / Liquid Samples
- Dilute 10x Extraction Buffer to 1x concentrated Buffer with PCR-grade water.
- Transfer 2 μl of the Blood/Liquid Sample into a tube containing 100 μl to 200 μl of 1x concentrated Extraction Buffer (a dilution of Blood 1:50 to 1:100 in 1x Extraction Buffer is recommended).
- Close the tube and vortex for 15 sec
- Incubate the tube at room temperature (20-25 °C) for 2-3 min.
- Transfer 1-2 μl of the supernatant into a 20 μl qPCR assay or 2-5 μl into a 50 μl qPCR assay.
b) Samples from nasal or throat swabs
- Dilute 10x Extraction Buffer to 1x concentrated Buffer with PCR-grade water.
- Transfer 200 μl 1x Extraction Buffer into a 1.5 ml microtube
- Cut off the cotton tip with the collected nasal or throat swab and place it in the micro tube
- Close the tube and vortex for 15 sec
- Incubate at room temperature (20-25 °C) for 2-3 min
- Remove the cotton tip and squeeze it out at the rim of the tube
- Centrifuge briefly and transfer 1-2 μl of the supernatant into a 20 μl qPCR assay or 2-5 μl into a 50 μl qPCR assay.
c) Samples from Animal or Plant Tissue
- Dilute 10 x Extraction Buffer to 1 x concentrated Buffer with PCR-grade water.
- Prepare a small piece from animal or plant tissue not exceeding 8 mm in diameter
- Crack plant seeds to less than 1 mm in diameter using a BeadBeater, Tissue Lyser or small hammer
- Place the sample in a 1.5 ml microtube
- Add 1x concentrated Extraction Buffer to the tissue sample as following:
| Sample size (diameter) | 1-2 mm | 3-4 mm | 5-8 mm |
| 1x Extraction Buffer | 50 μl | 100 μl | 200 μl |
- Mix briefly by tapping or vortexing and make sure that the sample is soaked with Extraction Buffer
- Incubate at room temperature (20-25 °C) for 3 min
- Centrifuge briefly and transfer 1-2 μl of the supernatant into a 20 μl qPCR assay or 2-5 μl into a 50 μl qPCR assay.
Signal word: Danger
Hazard statements:
- H314 Causes severe skin burns and eye damage.
Precautionary statements:
- P280 Wear protective gloves/protective clothing/eye protection/face protection/hearing protection/....
- P301 + P330 + P331 IF SWALLOWED: rinse mouth. Do NOT induce vomiting.
- P303 + P361 + P353 IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water [or shower].
- P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
- P363 Wash contaminated clothing before reuse.
- P405 Store locked up.
