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BacReadyTM Multiplex PCR System
Description
BacReadyTM Multiplex PCR System is a powerful reagent kit for both rapid preparation and multiplex PCR amplification of bacteria genomic DNA and plasmid DNA. Genomic and plasmid DNA are directly released from bacteria (both gram-positive and gram-negative) cells in a single step by adding a proprietary reagent directly to bacteria samples without DNA purification. The genomic and plasmid DNA can then be used immediately in PCR amplification of multiple gene targets (up to >1,000) or stored at 4°C for future use (stable at least 6 months at 4°C).
Key features of genscript PCR systems:
- Easy to perform: very simple and rapid procedure to extract genomic DNA and plasmid DNA in a single step.
- High specificity: highly specific amplification of genomic DNA using "HotStart" ScriptTM DNA polymerase (a genscript proprietary DNA polymerase).
- Multiplex PCR: up to >1,000 DNA sequences can be amplified using multiplex PCR primers.
- Super sensitivity: a single bacteria cell can be used to amplify both genomic DNA and plasmid DNA.
Applications
For genomic DNA prepared from: gram-positive or gram-negative bacteria.
For applications such as:
- SNP genotyping and mutation detection
- Target detection in transgenic plant
- DNA sequencing and cloning
- Quantitative PCR
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Technical Manual:
Components:
MSDS: 20050711104519 (PDF) PROTOCOL: 20070410225700 (PDF) Application Examples:A. Gram-negative Bacteria Genomic DNA Preparation and PCR Amplification. To test genomic DNA preparation from gram-negative bacteria and PCR amplification, a pairs of primers specific for E. coli are selected to amplify part of 16S rRNA gene. The expected PCR product is 544 bp. And the sequences of the two primers used in the experiments are: Forward primer: 5’-GGAAGAAGCTTGCTTCTTTGCTGAC-3’ Reverse primer: 5’-AGCCCGGGGATTTCACATCTGACTTA-3’ Genomic DNAs were prepared from four different E. coli cultures and amplified using BacReadyTM Multiplex PCR system (with Enzyme) following the kit instructions. The results were shown in Figure 1.  Figure 1. PCR analysis of genomic DNA prepared from bacteria samples. Genomic DNAs were prepared from 4 different E. coli cultures and amplified using BacReady(TM) Multiplex PCR kit following the kit instructions. M is 100 bp DNA marker lane. Lane 1 and 2 are culture No. 1; Lane 3 and 4 are culture No. 2, and so on. Lane 9 is a negative control. PCR products are 544 bp. B. Gram-positive Bacteria Genomic DNA Preparation and PCR Amplification. To test genomic DNA preparation from gram-positive bacteria and PCR amplification, a pairs of primers specific for gram-positive bacteria are selected to amplify part of rRNA gene. The expected PCR product is 355 bp. And the sequences of the two primers used in the experiments are: Forward primer: 5’-GCGRCTCTCTGGTCTGTA-3’ Reverse primer: 5’-GACGACAGCCATGCASCACCTGT-3’ Genomic DNAs were prepared from five different samples and amplified using BacReadyTM Multiplex PCR system (with Enzyme) following the kit instructions. The results were shown in Figure 2.  Figure 2. Bacteria genomic DNA was prepared and amplified using BacReady(TM) Multiplex PCR System (with Enzyme). Five different samples were tested. Lane 1 and 2 were yogurt samples; Lane 3, 4 and 5 were bacteria samples picked from slants, they were Bacillus globigii, Bacillus ubtillus and Micrococcus leteus, respectively. Lane 6 is a negative control. PCR products are 355 bp. Figure 2 shows that all the five DNA samples prepared using the kit produced the expected PCR products. There is little difference between the PCR amplification from 5 samples. This demonstrates the high quality and reproducibility of BacReady(TM) PCR System. |