Spin column based genomic DNA purification from bacteria
For in vitro use only!
Shipping: shipped at ambient temperature
Storage Conditions: store at ambient temperature
Shelf Life: 12 months
Availability Restriction: Exclusively distributed in Japan by Greiner BioOne
The spin column based Bacteria DNA Preparation Kit is designed for rapid and high purity isolation of genomic DNA from Gram-positive and Gram-negative bacteria. The spin column based method completely removes PCR inhibitors such as divalent cations and proteins resulting in a high purity preparation of genomic DNA. There is no use of phenol or chloroform, handling is safe and does not produce any harmful waste.
Column based genomic DNA purification kits yield up to 30 μg DNA sized from 200 bp to 50 kb per preparation.
RNase A (before use, add double distilled water as indicated on the bottle) - store at -20 °C
Lysozyme (before use, add double distilled water as indicated on the bottle) - store at -20 °C
Proteinase K (before use, add double distilled water as indicated on the tube) - store at -20 °C
Washing Buffer (before use, add 96-99 % Ethanol as indicated on the tube)
Spin Columns and 2 ml Collection Tubes
To be provided by you:
96-99 % Ethanol
Double distilled water
Microtubes 1.5 or 2.0 ml
The obtained DNA is suitable for a variety of applications, including real-time PCR, southern blot analysis, genotyping and discovery or validation of SNP/SSR markers.
Before start, add the following components (not included in the kit) as indicated on the respective bottle/tube:
For XS pack (10 preps): Before start, add 100 μl dd-water to the Proteinase K tube, 25 μl dd-water to the Lysozyme tube, 30 μl dd-water to the RNase A tube and 9.6 ml 96-99 % Ethanol (not included in the kit) to the Washing Buffer bottle.
For S pack (50 preps): Before start, add 500 μl dd-water to the Proteinase K tube, 125 μl dd-water to the Lysozyme tube, 150 μl dd-water to the RNase A tube and 48 ml 96-99 % Ethanol (not included in the kit) to the Washing Buffer bottle.
For L pack (250 preps): Before start, add 500 μl dd-water to each Proteinase K tube, 125 μl dd-water to each Lysozyme tube, 150 μl dd-water to each RNase A tube and 120 ml 96-99 % Ethanol (not included in the kit) to each Washing Buffer bottle.
|Resuspension Buffer||3.2 ml||16 ml||80 ml|
|Lysis Buffer||3.2 ml||16 ml||80 ml|
|Binding Buffer||3.2 ml||16 ml||80 ml|
|1.5 mg||7.5 mg||5x 7.5 mg|
|2.5 mg||12.5 mg||5x 12.5 mg|
|1 mg||5 mg||5x 5 mg|
|Activation Buffer||1.2 ml||6 ml||30 ml|
|Washing Buffer||add 9.6 ml Ethanol |
(final volume 12 ml)
|add 48 ml Ethanol |
(final volume 60 ml)
|add 120 ml Ethanol to each bottle (final volume 150 ml each)|
|Elution Buffer||1 ml||5 ml||25 ml|
It is essential to use the correct amount of starting material in order to obtain optimal DNA yield and purify. A maximum amount of 108 bacteria cells can generally be processed. Overnight cultured bacteria cells can be processed. Cell pellets can be stored at -70 °C for several months.
1. Preparation from Gram-positive bacteria
A Cell Resuspension:
B Cell Lysis:
C Column Activation [optional]:
D Column Loading:
E Primary Washing:
F Secondary Washing:
G Elution of DNA:
2. Preparation from Gram-negative bacteria
A Cell Lysis:
B Column Activation [optional]:
C Column Loading:
D Primary Washing:
E Secondary Washing:
F Elution of DNA: