- 03-3545-5720（東京）
- 06-7739-7114（大阪）
MasterPure™ Gram Positive DNA Purification Kit
グラム陽性菌からのDNA抽出
MasterPure™ Gram Positive DNA Purification Kitはグラム陽性バクテリアからDNAを精製する全ての試薬を提供します。Ready-Lyse™とGram Positive Cell Lysis Solutionで処理することでバクテリアを溶解させることができます。
Ready-Lyse™ Lysozymeは、非哺乳類の組換えのリゾチームで、中性pHで高比活性をもち、かつネットチャージを持たない安定な溶液ですので、リゾチームを溶解させる必要もなく、DNAに結合しません。1,2
Applications
Fosmidライブラリー構築やPCR、制限酵素処理、サザンブロットなどのためのグラム陽性バクテリアからのゲノムDNA精製
Benefits
- PCR-ready DNA
- Scalable method
- High molecular weight DNA
- No extra enzyme to purchase
- Tested on Bacillus, Streptococcus and Listeria
Table 1. Incubation Times Needed for DNA Recovery from 1 ml of Gram Positive Bacterial Culture.
| Species | Culture Medium | Ready-Lyse™ Incubation | DNA Yield ug/ml |
| Bacillus subtilis | Brain-Heart Infusion (BHI) | 30 min | 9.0 |
| Listeria monocytogenes | BHI | Overnight | 3.3 |
| Staphylococcus aureus | BHI | Not needed | 8.0 |
| Streptococcus mutans | Todd-Hewitt | Overnight | 3.0 |
| Lactococcus lactis | M17 | 30 min | 1.1 |
Figure 1
B. subtilis DNA
Figure 2
PCR of BacF/BacR
Figure 1. Electrophoretic Analysis of DNA Purified Using the MasterPure™ Gram Positive DNA Purification Kit.
A.DNA from Bacillus subtilis (ATCC 6051) was separated on a 1% agarose gel and stained with SYBRR Gold.Lane 1, kilobase ladder. Lane 2, 300 ng of B. subtilis DNA. B. Pulsed field gel electrophoresis of Bacillus subtilis (ATCC 6051) DNA. Lane 1, Phage lambda ladder. Lane 2, 300 ng of DNA. Lane 3, 600 ng of DNA.
Figure 2. PCR Product from DNA Purified Using the MasterPure™ Gram Positive DNA Purification Kit.
B. subtilis DNA (1 ul containing 0.5 ng) was amplified by PCR using forward primer 5'-AGG GTC ATT GGA AAC TGG G and reverse primer 5'-CGT GTT GTA GCC CAG GTC ATA. The cycle conditions were 95°C for 2 min and then 30 cycles of [95° C for 45 sec; 55° C for 45 sec; 72° C for 45 sec] followed by 72° C for 2 min. Lane 1, 100 bp ladder. Lane 2, PCR product from 0.5 ng of B. subtilis DNA. Lane 3, no template PCR.
References
- Hoffman, L.M. and B.W. Jarvis, (2003) EPICENTRE Forum. 10 (3), 3.
- Jarvis, B.W. and L.M. Hoffman, (2004) EPICENTRE Forum. 11 (3), 7.