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Bacterial Genomic DNA Isolation Kit

Isolates highly pure, intact bacterial DNA
Catalog #: K309
$285.00

Product Details

Cat # +Size K309-100
Size 100 isolations
Species Reactivity Bacterial
Applications Useful for PCR, Cloning, DNA hybridization, Southern Blotting
Features & Benefits • Simple & rapid method to isolate highly pure, intact DNA from bacteria
Kit Components • Buffer A [Re-suspension Buffer]
• Enzyme Mix A
• RNAse A
• Buffer B [Reaction Buffer]
• Enzyme Mix B
• Buffer C [Binding Buffer]
• Buffer D [Wash Buffer]
• Buffer E [Elution Buffer]
• Spin Columns/Collection Tubes
Storage Conditions -20°C/RT
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Bacteria are one of the most abundant and diverse organisms on the planet, which take part in numerous critical ecosystem processes. Many bacterial species are pathogens that are responsible for causing a variety of human and animal diseases. In addition to their medical and ecological importance, bacteria are also used in various industrial applications such as production of enzymes and biofuels. BioVision’s bacterial genomic DNA isolation kit provides convenient and simple step-by-step method for isolating quality genomic DNA from gram-negative and gram-positive bacterial species. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column in the presence of high salt concentration, eliminating the use of toxic organic compounds or solvents. DNA purified by this kit is suitable for various downstream molecular biology applications such as PCR, cloning, DNA hybridization, and Southern Blotting


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Why is my genomic DNA sheared?
1.The genomic DNA was handled improperly - Pipetting steps should be handled as gently as possible. Reduce vortexing times during mixing steps (no more than 10-15 seconds).
2.The cells are old - Older cultures contain prematurely lysed cells which release endonucleases and can degrade DNA. Fresh cultures are recommended.
Why is my yield of genomic DNA low?
1. The sample is old/overgrown - The culture may have been overgrown, allowing lysis of older cells to occur more readily. This will lead to premature degredation of the genomic DNA. It may be necessary to use bacterial cultures before they reach maximum density.
2. Incomplete lysis of cells - Extend the incubation time of Proteinase K digestion or reduce the amount of bacterial cells used for lysis. Increase the lysozyme incubation time for gram positive strains.
3. The DNA elution is incomplete - Ensure that centrifugation at 14,000 x g is performed after the 3,000 x g centrifugation cycle, to ensure that all the DNA is eluted.
Why is the lysate very gelatinous prior to loading onto the column?
1. The lysate/binding solution mixture is not homogeneous - To ensure a homogeneous solution, vortex for 10-15 seconds before applying the lysate to the spin column.
2. The sample is too large - Too many cells are in the lysate preparation. Ensure that the amount of cells used is less than of 2 x 109 viable cells, and that no more than 1 mL of culture is applied to the column.
Why is I my spin column getting clogged?
Too many cells were applied to the column. Ensure that the amount of cells used is less than 2 x 109 viable cells, and that no more than 1 mL of culture is applied to the column. Clogging can be alleviated by increasing the g-force and/or centrifuging for a longer period of time until the lysate passes through the column.
Can this kit work with Thermophilic bacteria?
This kit employs a general technique to isolate genomic DNA from most of the bacterial species including Thermophilic bacteria.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.