Over the past several decades, mitochondrial DNA (mtDNA) has played an increasingly important role in forensic analyses of various criminal cases. A few hairs left at a crime scene contain enough mtDNA for extraction. The hair shaft, which protrudes out of the scalp, does not contain any nuclear DNA. It does, however, contain mtDNA. While nuclear DNA is present in only two copies per cell, the small circular mtDNA molecule is present in hundreds to thousands of copies per cell making it very abundant. Mitochondrial DNA is maternally inherited, and all of a woman’s offspring will have the same mtDNA profile. An advantage of this is that a single maternal relative of that person may provide a reference sample for comparison to a sample found at a crime scene.
Norgen’s Hair Mitochondrial DNA Isolation Kit provides a fast, reliable, and simple procedure for isolating mtDNA from hair shafts. Purification is based on spin column chromatography and the DNA is preferentially purified from other components. Typical yields will vary depending on the sample input volume used. The purified DNA is compatible with all downstream applications including PCR and NGS.
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Storage Conditions and Product Stability
Store DTT at -20°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers. The kit contains a ready to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
| Component | Cat. 69400 (50 preps) |
|---|---|
| DTT | 6 mL |
| Proteinase K | 4 mL |
| Lysis Additive A | 6 mL |
| Lysis Buffer B | 20 mL |
| Wash Solution A | 38 mL |
| Elution Buffer B | 8 mL |
| Micro Spin Columns | 50 |
| Elution Tubes | 50 |
| Collection Tubes | 50 |
| Product Insert | 1 |
Magnetic Beads (tRNA Purification)
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
The Magnetic Beads (tRNA Purification) solves the tRNA and short DNA/RNA purification problem. The beads with our proprietary technology purify tRNA effectively by removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants. The magnetic bead reagents can be used for oligo (>70 nt) applications.
Workflow without large RNA/DNA contamination
In the case of samples contaminated with RNA/DNA such as rRNA and DNA, our magnetic beads can effectively remove RNA/DNA that are 180 nt and larger. Purified tRNA are ideal for applications requiring high quality, as the fragments are free of impurities and contaminants.
Workflow with large RNA/DNA contamination
Comparison of tRNA and 70 nt oligos recovery. BioDynami beads (tRNA purification) successfully recovered DNA fragments both yeast tRNA and 70 nt oligos.
Recovery of tRNA and 70 nt oligos with BioDynami magnetic Beads (tRNA Purification). Yeast tRNA and 70 nt oligos were used as input. Input and recovered oligos were quantified with ssDNA Quantification kit (BioDynami Cat. # 40043) and RNA Quantification kit (BioDynami Cat. # 40044).
Depletion of larger RNAs. Left panel: depletion of 28S rRNA and 18S rRNA. Right panel: depletion of RNA of 180 nt and larger.
Features
Removal of unwanted components and other impurities
Purification of tRNA and oligos (>70 nt)
tRNA
RNA fragments 70 nt or longer
DNA/RNA hybrid fragments 70 nt or longer
Oligo and chimeric oligo 70 nt or longer
dsDNA fragments 70 bp or longer
ssDNA fragments 70 nt or longer
Removal of larger RNA/DNA contamination:
18S rRNA
28S rRNA
RNA/DNA> 180 nt
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
For selective enrichment culture of Listeria monocytogenes in food.
Tryptone, proteose peptone, meat extract, yeast paste powder provide nitrogen sources, vitamins, amino acids, and growth factors; sodium chloride can maintain a balanced osmotic pressure; sodium dihydrogen phosphate and potassium dihydrogen phosphate as buffering agents; esculin are fermentable sugars; lithium chloride and other antibiotics can inhibit Gram negative bacteria and most Gram positive bacteria.
| Ingredients | /liter |
| Enzymatic digest of animal tissues ( Proteose peptone) | 5.0g |
| Enzymatic digest of casein ( Tryptone) | 5.0g |
| Meat extract | 5.0g |
| Yeast extract | 5.0g |
| Sodium chloride | 20.0g |
| Disodium hydrogen phosphate dihydrate | 12.0g |
| Potassium dihydrogen phosphate | 1.35g |
| Aesculin | 1.0g |
| Lithium chloride | 3.0g |
| pH7.2±0.2 at 25°C | |
Suspend 57.4g in 1 L of distilled or deionized water. Heat with frequent agitation and boil to completely dissolve the powder. Distribute into flasks. Autoclave at 121℃ for 15 minutes. Cool to 45-50℃,aseptically add the contents of 1 vial of Fraser Selective Supplement (SR0120) to each 225 mL of base to prepare FB1 solution; Add one reagent (SR0130) to each100 mL of FB2 culture medium to prepare FB2 enrichment solution.
Cultural characteristics observed after incubation at 35-37°C for 46~50 hours
| Quality control strains | Growth | Characteristics |
| Listeria monocytogenes ATCC19115 + Escherichia coli ATCC25922 + Enterococcus faecalis ATCC29212 | >20 cfu On PALCAM | Gray to black colony count with black halo |
| Escherichia coli ATCC25922 | < 100 colonies on TSA | Inhibition |
| Enterococcus faecalis ATCC29212 |
2-30℃，Keep container tightly closed, avoid direct sunlight.
Use before expiry date on the label.
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
On June 14, 2024
ISO 11290
Intended Use For selective enrichment culture of Listeria monocytogenes in food. Principle and Interpretation Tryptone, proteose peptone, meat extract, yeast paste powder provide nitrogen ……