Milk Bacterial DNA Isolation Kit (Cat. 21550)50 preps
Format: Spin Column
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Milk Bacterial DNA Isolation Kit
Genomic DNA can be isolated from as few as 10 bacterial cells in 1 mL of milk
Isolate genomic DNA from both Gram-negative and Gram-positive bacteria in milk
Can process challenging samples such as mastitic milk
Inhibitor-free DNA is ready for PCR, qPCR, Southern Blot, sequencing & more
Fast and efficient spin-column format
This kit provides a rapid spin column method for the isolation and purification of genomic DNA from both Gram-negative and Gram-positive bacteria in milk samples. It can also be used to process challenging milk samples such as Mastitic milk for example. The kit is highly sensitive and can isolate DNA from a cell density of as little as 10 bacteria contained in 1 mL of milk. Genomic DNA is isolated in 45 minutes, free of inhibitors and ready for any number of downstream applications including PCR, qPCR and Southern Blot analysis, sequencing and more.
Figure 1. Isolation and Detection of DNA from as Little as 10 Bacterial Cells in 1 mL of Milk. Increasing amounts of E coli (101cells, 103cells, 105cells, 107cells) were added to 1 mL pasteurized milk samples, and the bacterial DNA was subsequently isolated using Norgen's Milk Bacterial DNA Isolation Kit with a 200 µL elution volume. One microliter of the isolated genomic DNA was then detected in a 20 µL real-time PCR reactionusing primers specific to E coli. Bacterial genomic DNA could be isolated and detected from all the 1 mL milk samples, including the sample that contained only 10 bacterial cells (blue line).
Figure 2. End-Point PCR of Milk Bacterial DNA samples.One milliliter of milk was spiked with various amount of E coli. Bacterial DNA was isolated in duplicate using Norgen's Milk Bacterial DNA Isolation Kit, and 1 µL of each 200 µLelution was used for conventional PCR. For analysis 10 µL of each PCR was loaded on an agarose gel. The amount of bacteria in each milk sample is as follows: A = 107cells, B = 105cells, C = 103cells, D = 101cells. Lane X is a negative control (no template) and Lane M is Norgen's PCRSizer 100bp DNA Ladder.
Figure 3. Hierarchial Clustering Dendrogram of Milk Related Microorganisms Found in Milk Samples.DNA was isolated using Norgens Milk Bacterial DNA Isolation Kit and the 16S Metagenomic Sequencing Library was prepared according to the Illumina MiSeq System. This dendogram shows a hierarchial clustering of samples based on genus-level classifications. The barchart beneath each sample shows the relative abundance of its genus-level classifications.
*The range of the DNA yield will vary depending upon a number of factors including bacterial species and type of milk (fat content and %)
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The Lysozyme should be stored at -20°C upon arrival, and the Resuspension Solution A should be stored at -20°C after addition of the lysozyme. The lyophilized Proteinase K should be stored at -20°C upon arrival and after reconstitution. This kit is stable for 1 year from the date of shipment.