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By combining Proteinase K lysis with the speed and convenience of silica membrane purification, ISOLATE II Genomic DNA Kit provides a fast method for the purification of high-quality genomic DNA from a variety of starting materials.
By combining Proteinase K lysis with the speed and ease-of-use of silica-membrane purification, the ISOLATE II Genomic DNA Kit provides a fast method for the purification of high-quality genomic DNA from a variety of starting materials, such as mouse or rat tails, bacteria, yeast, dried blood spots, genomic/viral DNA from blood, hair follicles, paraffin-embedded issue (FFPE), insects, dental swabs, buccal swabs, stool viruses (e.g. CMV), Mycobacterium tuberculosis or Legionella pneumophila in sputum or bronchoalveolar lavage, EHEC bacteria in food, bacterial DNA (e.g. Chlamydia trachomatis) from cultures, biological fluids or clinical specimens, bacterial DNA (e.g. Borrelia burgordferi) and viral DNA (e.g. CMV) from urine.
ISOLATE II Genomic DNA Kit has been designed to deliver optimal performance in qPCR in tandem with SensiFAST Real-Time PCR Kits and in end-point PCR with MyTaq DNA Polymerase.
Excellent results from a variety of samples
Genomic DNA was isolated in duplicate from E. coli, mouse tail, mouse lung, HeLa cells and 3T3 cells, using ISOLATE II Genomic DNA Kit (lanes 1-5 and 6-10 respectively) and used to amplify a fragment of the Rn 18s gene. The results illustrate the consistency between replicates and between completely different species.
Consistency between species
Genomic DNA was isolated from hair samples of eastern lowland gorilla, western gorilla, northern plains gray langur, white-headed langur and red-shanked douc (lanes 1-5 respectively). Genomic DNA was isolated using ISOLATE II Genomic DNA Kit and a fragment amplified to illustrate the consistency in quality, purity and yield of the DNA extracted.
Reagent |
10 Preps |
50 Preps |
250 Preps |
ISOLATE II Genomic DNA Spin Columns |
10 |
50 |
250 |
Collection Tubes (2 mL) |
20 |
100 |
500 |
Lysis Buffer GL |
5 mL |
20 mL |
100 mL |
Buffer G3 |
10 mL |
15 mL |
75 mL |
Wash Buffer GW1 |
6 mL |
30 mL |
150 mL |
Wash Buffer GW2 (concentrate) |
6 mL |
12 mL |
50 mL |
Elution Buffer G |
13 mL |
13 mL |
60 mL |
Proteinase K (lyophilized) |
6 mg |
30 mg |
2 x 75 mg |
Proteinase K Buffer PR |
1.8 mL |
1.8 mL |
8 mL |
Bench Protocol Sheet |
1 |
1 |
1 |
All components should be stored dry and at room temperature.
When stored under the recommended conditions and handled correctly, full activity of reagents is retained until the expiry date indicated on the outer box label.
Ambient temperature.
For the optimal recovery of large RNA/DNA fragments it is necessary to optimize the elution step. To increase the recovery rate it would be helpful to use a larger volume for elution, incubate the column with the elution buffer prior to centrifugation and use repeated elution steps. To avoid excessive dilution it may be helpful to reapply the eluted fraction again. Furthermore it could be helpful for DNA elution to heat the elution buffer to 70°C.