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LAMP Green™ *50X DMSO Solution*

LAMP detection of BRCA1 in HeLa cells. 500 ng (Green), 50 ng (Blue), 5 ng (Orange), and NTC (Black) of gDNA in HeLa cells was used in LAMP reaction with LAMP Green™ fluorescent dye using ABI 7500 qPCR machine.
LAMP detection of BRCA1 in HeLa cells. 500 ng (Green), 50 ng (Blue), 5 ng (Orange), and NTC (Black) of gDNA in HeLa cells was used in LAMP reaction with LAMP Green™ fluorescent dye using ABI 7500 qPCR machine.
LAMP detection of BRCA1 in HeLa cells. 500 ng (Green), 50 ng (Blue), 5 ng (Orange), and NTC (Black) of gDNA in HeLa cells was used in LAMP reaction with LAMP Green™ fluorescent dye using ABI 7500 qPCR machine.
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Physical properties
Molecular weight578.73
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


Molecular weight
578.73
Isothermal amplification methods provide detection of nucleic acid target sequence in a streamlined, exponential manner. The existing dyes that successfully detect the LAMP reaction are mostly based on the end point method using the colorimetric analysis. LAMP Green™ enables real-time fluorescence measurement of a LAMP reaction. The dye can be detected using the SYBR or FAM channel on common real-time PCR instruments. LAMP Green™ is compatible with the agarose gel electrophoresis, making it possible to run on a gel and analyze it by gel imager.

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

Table 1. 25 µL LAMP reaction mix.
ComponentsDNA/RNA target detectionNo Template Control (NTC)
LAMP master mix (2X)12.5 µL12.5 µL
LAMP primer mix (10X)2.5 µL2.5 µL
LAMP Green™ (50X)0.5 µL0.5 µL
Target DNA/RNAVaries0 µL
ddH2OVaries9.5 µL
Total Volume25 µL25 µL
The following protocol can be used as a guideline and should be optimized according to your experimental needs.
  1. Thaw all components to be used at room temperature and place on ice. Vortex briefly to mix and centrifuge to collect material.
  2. Prepare reaction mix as described in Table 1. Volumes are listed for 25 µL/LAMP reaction. Adjust volume accordingly as per use. If necessary, LAMP Green™ concentration can be optimized.
  3. Vortex reaction mix and centrifuge to collect material.
  4. Seal the reaction vessel.
  5. Incubate at 65°C for 30 minutes. If necessary, time can be extended (i.e., for low copy targets, challenging sample types, or reactions known to produce slower amplification times).
  6. If using with a qPCR machine, the signal can be read with either a SYBR or FAM filter in real-time. For microplate reader-based assays, measure the signal at Ex/Em = 490/525 nm (Cutoff = 515 nm).
  7. If reaction products will be analyzed after LAMP reaction is completed, then deactivate the enzyme by heating at >80°C for 5 minutes.
    Note     Deactivation can be performed for LAMP master mix provided by the manufacturer. LAMP Green™ is compatible with agarose gel electrophoresis, so samples can be analyzed on an agarose gel.
     

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of LAMP Green™ *50X DMSO Solution* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM172.792 µL863.961 µL1.728 mL8.64 mL17.279 mL
5 mM34.558 µL172.792 µL345.584 µL1.728 mL3.456 mL
10 mM17.279 µL86.396 µL172.792 µL863.961 µL1.728 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
/=x=

Images


References


View all 16 references: Citation Explorer
Comparison of COVID-19 laboratory diagnosis by commercial kits: Effectivity of RT-PCR to the RT-LAMP.
Authors: Artik, Yakup and Coşğun, Alp B and Cesur, Nevra P and Hızel, Nedret and Uyar, Yavuz and Sur, Haydar and Ayan, Alp
Journal: Journal of medical virology (2022): 1998-2007
SARS-CoV-2's high rate of genetic mutation under immune selective pressure: from oropharyngeal B.1.1.7 to intrapulmonary B.1.533 in a vaccinated patient.
Authors: Musso, Nicolò and Maugeri, Jessica Giuseppina and Bongiorno, Dafne and Stracquadanio, Stefano and Bartoloni, Giovanni and Stefani, Stefania and Di Stefano, Emanuela Daniela
Journal: International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases (2022): 169-172
Rapid quantification of fecal indicator bacteria in water using the most probable number - loop-mediated isothermal amplification (MPN-LAMP) approach on a polymethyl methacrylate (PMMA) microchip.
Authors: Fu, Jing and Chiang, Elaine Li Ching and Medriano, Carl Angelo Dulatre and Li, Liyan and Bae, Sungwoo
Journal: Water research (2021): 117172
Severe long-delayed malaria caused by Plasmodium malariae in an elderly French patient.
Authors: Marteau, Anthony and Ouedraogo, Elise and Van der Meersch, Guillaume and Akhoundi, Mohammad and Souhail, Berenice and Cohen, Yves and Bouchaud, Olivier and Izri, Arezki
Journal: Malaria journal (2021): 337
Confirmed validation of an innovative PCR-assay without DNA extraction for multiplex diagnosis of factor V Leiden and prothrombin gene variants.
Authors: Desjonquères, A and Ménard, A and Detemmerman, L and Ternisien, C and Fouassier, M and Gillet, B and Béné, M C and Le Bris, Y
Journal: Thrombosis research (2019): 143-145
[Clinical Application of Loop-mediated Isothermal Amplification in Detection of Mycoplasma Pneumoniae].
Authors: Yan, Chun Xia and Lu, Wei Hong and He, Guo Chan and Wen, Ren Qing and Qian, Ying
Journal: Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae (2019): 203-207
Rapid and in-situ detection of fecal indicator bacteria in water using simple DNA extraction and portable loop-mediated isothermal amplification (LAMP) PCR methods.
Authors: Lee, Seunguk and Khoo, Valerie Si Ling and Medriano, Carl Angelo Dulatre and Lee, Taewoo and Park, Sung-Yong and Bae, Sungwoo
Journal: Water research (2019): 371-379
Cloning of BBTV (Banana Bunchy Top Virus) components and screening of BBTV using functionalized gold nanoparticles.
Authors: Kumar, P and Arun, V and Lokeswari, T S
Journal: 3 Biotech (2017): 225
Genetic variability and discrimination of low doses of Toxocara spp. from public areas soil inferred by loop-mediated isothermal amplification assay as a field-friendly molecular tool.
Authors: Ozlati, Maryam and Spotin, Adel and Shahbazi, Abbas and Mahami-Oskouei, Mahmoud and Hazratian, Teimour and Adibpor, Mohammad and Ahmadpour, Ehsan and Dolatkhah, Afsaneh and Khoshakhlagh, Paria
Journal: Veterinary world (2016): 1471-1477
LAMP-PCR detection of ochratoxigenic Aspergillus species collected from peanut kernel.
Authors: Al-Sheikh, H M
Journal: Genetics and molecular research : GMR (2015): 634-44