Magigen MagicScript 2xLAMP Amplification Mix with dye
Catalog
F001L
Intended use
Magigen MagicScript LAMP kit is one-step solution for Loop-Mediated Isothermal Amplification(LAMP) of DNA targets. The kit contains fluorescent dye, dNTPs, metal ion indicators for LAMP reaction.
Size
500T
Storage conditions and Expiration
Storage conditions: -20℃,avoid light.
Expiration: 12 months.
Contents of the kit
Item
F001S
F001L
2×LAMP Amplification Mix
625μLx1Tube
625μLx10Tube
Detection principle
LAMP, Loop-mediated isothermal amplification, uses 4 primers recognizing 6 distinct regions of target DNA fora highly specific amplification reaction. This method employs a strand-displacing DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. The method provides high amplification efficiency of specific DNA, up to 10fold, at a constant temperature (about 60-65 °C) in 15-60 Min. it is simple, high specificity, efficiency and rapidity. The products would enable their simple, easy and selective detection.
Applicable instrument
You can use a fluorescence quantitative PCR instruments, like Slan-96P,Strongmed, Shanghai, or MA-1600Q series Real-Time Quantitative Thermal Cycler.
Magigen LAMP kit DATA
Ultra sensitive
The sensitive is up to 10 copies/reaction when using the Magigen LAMP Kit to test the African Swine Fever Virus (ASFV) sample. It is stable at 100 copies/reaction.
Fig1. From left to right are the amplification curves of 10,000, 100 and 10 copies respectively.
Good stability
The nucleic acid amplification results showed that the performance remained unchanged after the kit repeated freezing and thawing for 1, 2, 3, 4, 5 times .
Operation procedure
1. DNA extraction.
You can use commercial DNA extraction kits to do DNA extraction. Spin column and magnetic bead method are recommended.
2. Methods
2.1 Take the components out, melt slowly on ice or at 4℃, mix well, centrifuge briefly, and set aside.
2.2 Make reaction mix in a centrifuge tube with the components in the list below.
Component
Recommended volume(μL)
2×LAMP Amplification Mix
12.5
F3/B3 Primers(10μM)
2
FIP/BIP Primers(10μM)
4
LF/LB(10μM)(opt)
2
Template
2
RNase-free Water
Up to 25
Vortex immediately after preparation and spin down gently, mix well, centrifuge briefly, put it into the instrument immediately to react.
2.3 Put the tube into the fluorescence quantitative PCR instruments with FAM channel, set the temperature to 60-65℃ (65℃ is recommended), reaction time30-60 min. The nonspecific amplification will increase significantly when reaction time beyond 50 min. Collect the fluorescent data every 60 second.
Note:
a. Generally, the reaction time is set to 45 min. The reaction time could be prolonged to 60 min if Loop primers are not used.
b. The primer combination and volume can be optimized and adjusted according to the actual situation.
