GenElute™ Blood Genomic DNA Kit
SIGMA/NA2020 - sufficient for 350 purifications
Synonym: Blood Genomic DNA; Gen Elute
Product Type: Chemical
Genomic DNA purified by GenElute™ Blood Genomic DNA kits is suitable for restriction enzyme digestions.
Restriction Enzymes, EcoR I and Hind III were used to digest genomic DNA isolated with GenElute™ Blood Genomic DNA kit. Whole blood was collected in 3 different anticoagulants: EDTA, heparin, and sodium citrate. A 100 ng aliquot of genomic DNA from each anticoagulant was initially digested with EcoR I (5 units per 1 μL digested at 37 °C for 1.5 hours) and Hind III (10 units per 1 μL digested at 37 °C for 1.5 hours) followed by electrophoresis (50 ng/lane) on a 0.8% agarose gel. Ladder (M) used was Lambda Hind III (Cat. No. D9780).
Restriction Enzymes, EcoR I and Hind III were used to digest genomic DNA isolated with GenElute™ Blood Genomic DNA kit. Whole blood was collected in 3 different anticoagulants: EDTA, heparin, and sodium citrate. A 100 ng aliquot of genomic DNA from each anticoagulant was initially digested with EcoR I (5 units per 1 μL digested at 37 °C for 1.5 hours) and Hind III (10 units per 1 μL digested at 37 °C for 1.5 hours) followed by electrophoresis (50 ng/lane) on a 0.8% agarose gel. Ladder (M) used was Lambda Hind III (Cat. No. D9780).
Whole Blood collected in three different anticoagulants was isolated with the GenElute™ Blood Genomic DNA kit and Supplier Q to obtain genomic DNA.
A. Purified genomic DNA from whole blood collected in vacutainer tubes, each containing a different anticoagulant (EDTA, heparin or sodium citrate). Samples were isolated with either the GenElute™ Blood Genomic DNA kit or Supplier Q following both protocols in detail. The genomic DNA (100 ng/lane) was analyzed on a 0.8% agarose gel to show overall comparability with Supplier Q. The whole blood (200 μL per sample) used was obtained from a human donor. Marker (M) used was Lambda Hind III (Cat. No. D9780).
B. Yields of genomic DNA compared with Supplier Q from 3 different anticoagulants. The amount of DNA was determined measuring absorbance at (A260/A280).
A. Purified genomic DNA from whole blood collected in vacutainer tubes, each containing a different anticoagulant (EDTA, heparin or sodium citrate). Samples were isolated with either the GenElute™ Blood Genomic DNA kit or Supplier Q following both protocols in detail. The genomic DNA (100 ng/lane) was analyzed on a 0.8% agarose gel to show overall comparability with Supplier Q. The whole blood (200 μL per sample) used was obtained from a human donor. Marker (M) used was Lambda Hind III (Cat. No. D9780).
B. Yields of genomic DNA compared with Supplier Q from 3 different anticoagulants. The amount of DNA was determined measuring absorbance at (A260/A280).
PCR amplification of a 388 bp product isolated from Genomic DNA.
Whole blood collected in 3 different anticoagulants was purified using the GenElute™ Blood Genomic DNA Kit or a comparable kit from Supplier Q. A 5 μL aliquot of each eluate was used as template in a 20 μL PCR reaction for 35 cycles. A 5 μL aliquot of each PCR reaction was resolved on a 2% agarose gel. The PCR marker (M) (Cat. No. P9577) used ranged from 50 bp to 2 kb.
Whole blood collected in 3 different anticoagulants was purified using the GenElute™ Blood Genomic DNA Kit or a comparable kit from Supplier Q. A 5 μL aliquot of each eluate was used as template in a 20 μL PCR reaction for 35 cycles. A 5 μL aliquot of each PCR reaction was resolved on a 2% agarose gel. The PCR marker (M) (Cat. No. P9577) used ranged from 50 bp to 2 kb.
| Quality Level | 200  |
| storage temp. | 15-25°C |
| room temp | |
| technique(s) | DNA purification: suitable |
| usage | sufficient for 350 purifications |
| Application: | The purified genomic DNA is ready for downstream applications such as: • restriction endonuclease digestions • PCR • Southern blots • sequencing reactions • cloning |
| Biochem/physiol Actions: | The starting material is lysed in a chaotropic salt-containing solution to ensure the thorough denaturation of macromolecules. The addition of ethanol causes the DNA to bind when the lysate is spun through a silica membrane in a microcentrifuge tube. A Prewash Solution is provided to help remove contaminants that are associated with aged (older than 24 hours) whole blood samples. After washing to remove contaminants, the DNA is eluted in 200 mL of a Tris-EDTA solution. The expected yields of genomic DNA will vary depending on the amount and nature of the starting material used (for example, 4 to 10 μg of RNase A-treated DNA can be isolated from 200 μl of fresh whole blood in less than one hour). DNA purified with this kit has an A260/A280 ratio between 1.6 and 1.9 and can be up to 50 kb in length. |
| Features and Benefits: | • Starting material: Up to 200 μl of fresh or aged blood • Expected yield: Up to 10 μg • Elution volume: 400 μl • Time required: <40 min • A260/A280 ratio: 1.6 - 1.9 • Compatible with many anticoagulants, including EDTA, Heparin, and Sodium Citrate |
| Features and Benefits: | • Typical yields 4-10 μg from 200 μL of whole blood • Pure genomic DNA with a A260/A280 ratio between 1.6 and 1.9 • Isolate DNA up to 50 kb in length |
| General description: | The GenElute Blood Genomic DNA kit provides a simple and convenient way to isolate pure genomic DNA from fresh or aged (older than 24 hours) whole blood. The kit combines the advantages of silica binding with a microspin format, and eliminates the need for expensive resins, alcohol precipitation, and hazardous organic compounds such as phenol and chloroform. |
| Legal Information: | GenElute is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | For additional information, please see www.sigma-aldrich.com/gen. |
| Symbol |     GHS05,GHS07,GHS08,GHS09 |
| Signal word | Danger |
| Hazard statements | H290 - H302 - H315 - H318 - H334 - H410 |
| Precautionary statements | P234 - P273 - P280 - P301 + P312 - P302 + P352 - P305 + P351 + P338 |
| Hazard Codes | Xn |
| Risk Statements | 22-36/37/38-42-52/53 |
| Safety Statements | 23-26-45-61 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 15-25°C; room temp |
| UNSPSC | 41105501 |
| Components | Proteinase K from Tritirachium album, lyophilized powder, Molecular Biology, BioUltra, ≥30 units/mg protein 2; Column Preparation Solution; RNase A solution |