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.

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Physical properties

Molecular weight	578.73
Solvent	DMSO

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure

Overview SDS Protocol

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.

Components	DNA/RNA target detection	No Template Control (NTC)
LAMP master mix (2X)	12.5 µL	12.5 µL
LAMP primer mix (10X)	2.5 µL	2.5 µL
LAMP Green™ (50X)	0.5 µL	0.5 µL
Target DNA/RNA	Varies	0 µL
ddH₂O	Varies	9.5 µL
Total Volume	25 µL	25 µL

The following protocol can be used as a guideline and should be optimized according to your experimental needs.

Thaw all components to be used at room temperature and place on ice. Vortex briefly to mix and centrifuge to collect material.
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.
Vortex reaction mix and centrifuge to collect material.
Seal the reaction vessel.
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).
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).
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 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	172.792 µL	863.961 µL	1.728 mL	8.64 mL	17.279 mL
5 mM	34.558 µL	172.792 µL	345.584 µL	1.728 mL	3.456 mL
10 mM	17.279 µL	86.396 µL	172.792 µL	863.961 µL	1.728 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Product Family

Name	Excitation (nm)	Emission (nm)
Indocyanine Green CAS 3599-32-4	789	814
Helixyte™ Green 20X Aqueous PCR Solution	498	522
Helixyte™ Green 10,000X Aqueous PCR Solution	498	522
Thiolite™ Green	510	525
CytoTell™ Green	510	525
LysoBrite™ Green	501	510
Q4ever™ Green 1250X DMSO Solution	503	527
Droplite™ Green	421	521

Images

Figure 1. 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.

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