MagBind Bacterial Genomic DNA Extraction Kit

 

Product Name	Cat. No.	Spec.
MagBind Bacterial Genomic DNA Extraction Kit	G3603-50T	50 T

 

 

The MagBind Bacterial Genomic DNA Extraction kit can rapidly release bacterial genomic DNA through the specially optimized bacterial lysis buffer system. Combined with the superparamagnetic magnetic beads, which are specially developed for nucleic acid extraction by our company, the genomic DNA in the lysate is selectively adsorbed under the action of the binding solution, and the small amount of impurities adsorbed by the magnetic beads is removed through the cleaning of the rinsing solution, and the adsorbed genomic DNA is released under the action of the elution solution. This kit is suitable for a variety of Gram-negative bacteria and most Gram-positive bacteria. The obtained high purity and high integrity genomic DNA can be directly used for subsequent PCR amplification, Southern hybridization, library preparation and other molecular biology experiments.

 

 

Lysozyme,Proteinase K and RNase A should be shipped with wet ice, and stored at -20℃; Other reagents are shipped and stored at room temperature for up to 12 months.

 

 

Component Number	Component	G3603-50T
G3603-1	Lysozyme	1 mL
G3603-2	Lysozyme Buffer	7 mL
G3603-3	Buffer MB1	12 mL
G3603-4	Buffer MB2	12 mL
G3603-5	Proteinase K	1 mL
G3603-6	RNase A	500 uL
G3603-7	SweMag Beads	1 mL
G3603-8	Buffer MBP	12 mL (18 mL of anhydrous ethanol added before use)
G3603-9	Buffer PW	24 mL (56 mL of anhydrous ethanol added before use)
G3603-10	Buffer TE	10 mL
Manual		One copy

 

 

1. If there is any precipitation of Buffer MB1, please dissolve it by heating at 65℃ and use it after it returns to room temperature.

2. Add 18 mL of anhydrous ethanol to Buffer MBP and 56 mL of anhydrous ethanol to Buffer PW before use.

3. Magnetic stands are required but not supplied in this kit.

 

 

1. Centrifuge 1-5 mL of overnight fresh bacterial culture at 12,000 rpm for 1 minute and discard the supernatant.

Note: For Gram-positive bacteria, lysozyme solution should be added to break the cell wall after collecting the organisms. The specific method is as following: add 130 µL Lysozyme Buffer, 20 µL Lysozyme, vortex and shake until the bacteria are thoroughly suspended, water bath at 37℃ for 60 min, mixing every 10 min with inversion.

2. Add 200 μL of Buffer MB1 to the centrifuge tube and mix by vortexing.

3. Add 20 μL of Proteinase K and 10 μL of RNase A to the centrifuge tube, mix well by pipetting or vortexing, and then incubate at 56°C for 20 min, at which time the solution appears clear (If the solution is not transparent at this point, extend the incubation time to 30 minutes, and mix once every 10 minutes).

4. add 200 uL of Buffer MB2 and 200 μL of absolute ethanol to the centrifuge tube and mix it upside and down, at which point a flocculent precipitate may appear.

5. Add 20 μL of SweMag Beads suspension to the centrifuge tube (SweMag Beads must be fully resuspended and evenly distributed prior to use), Blow with a pipette until the magnetic beads distribute evenly.

6. The magnetic beads were allowed to stand at room temperature for 10 min, and during the process, the magnetic beads were blown and mixed several times using a pipette to keep the magnetic beads in suspension.

7. Place the tube on a magnetic stand and stand for 30 seconds. When the supernatant is clear, aspirate and discard the supernatant.

8. Add 500 μL of Buffer MBP, remove the magnetic stand, pipet up and down gently until the magnetic beads are well dispersed. Place the tube on a magnetic stand for 30 seconds. When the supernatant is clear, aspirate and discard the clear supernatant.

9. Add 600 μL of Buffer PW, remove the magnetic stand, pipet up and down gently until the magnetic beads are well dispersed. Place the tube on a magnetic stand for 30 seconds. When the supernatant is clear, aspirate and discard the clear supernatant.

10. Repeat step 9.

11. Open the centrifuge tube lid, leave it at room temperature for 5-10 minutes to ensure complete evaporation of residual ethanol. (Do not allow them to dry out to avoid affecting the nucleic acid yield)

12. Remove the magnetic stand, add 50–100 μL Buffer TE or Nuclease-free Water and pipet up and down gently to resuspend the magnetic beads, stand at room temperature for 5 minutes.

13. Place the tube on a magnetic stand until the magnetic beads are fully adsorbed, then aspirate the supernatant into a new Nuclease-free centrifuge tube to obtain high-purity genomic DNA.

 

 

1. Be sure to read this product manual carefully before operation.

2. Fresh organisms should be used as much as possible to ensure the yield and integrity of the genomic DNA obtained by extraction.

3. The starting amount of extracted bacteriophage should never be too much and should be sufficiently lysed, otherwise the yield and purity of genomic DNA may be affected.

4. Magnetic bead suspensions should be stored to avoid freezing.

5. Magnetic beads are easy to settle, and should be shaken well before use to keep them in uniform suspension.

6. Before adding the magnetic beads, other reagents should be mixed well.

7. Do not dry the magnetic beads for a long time as this may affect the DNA elution efficiency.

 

For Research Use Only!