Description
LavaLAMP RNA Component Kit, 500 x 25 µL Reactions
Sensitive, fast RNA detection with this fully optimisable RNA LAMP (RT-LAMP) kit.
Both LavaLAMP RNA Component Kits contain: 10X LavaLAMP RNA Buffer, LavaLAMP RNA Enzyme, Magnesium Sulphate, 100 mM, RNA Positive Control LAMP Primer Mix and RNA Positive Control. The LavaLAMP RNA Component Kit with Dye also contains Green Fluorescent Dye for fluorescent detection of amplified DNA. The Green Fluorescent Dye is also available separately.
Key features
- Isothermal Amplification: Enables running amplification reactions using less complex and lower cost instruments.
- Lyophilisation-ready: All kit components are lyophilisation compatible, thus avoiding redesign of assays to remove components known to inhibit lyophilisation.
- Fully Optimisable: Non-master mix kit format gives you complete control of reaction formulation to maximise assay perfomance.
- Matched Performance to LavaLAMP RNA Master Mix: Assist current users of the master mix product by streamlining additional assay optimisation with this kit.
Product information
Like the related, LavaLAMP™ RNA Master Mix, this component kit is built around a proprietary DNA polymerase with strand displacement activity that works on both RNA and DNA templates. With these unique features, it is possible to perform RNA LAMP (loop-mediate isothermal amplification starting with RNA targets/templates) using a single DNA polymerase. This enzyme first copies the RNA into cDNA, and then loop mediated isothermal amplification takes over to amplify the initial RNA target sequence present in the new cDNA making amplified and detectable target-specific, dsDNA.
The LavaLAMP RNA Component Kit is a deconstruction of the LavaLAMP RNA Master Mix, and as such, when reactions are setup using the initial experimental reaction conditions outlined in the Component Kit User Manual, the composition of that reaction matches the composition of the recommended initial Master Mix reaction outline in its User Manual. The individual reagent format of this kit, as opposed to the master mix format, enables complete control of each LAMP assay allowing you to carefully identify the best reaction conditions for your specific target and primer set.
To aid in assay development and future lyophilisation of LavaLAMP RNA-enzyme based reactions, all components in this kit are compatible with lyophilisation. This feature means that reactions will not have to be re-formulated or re-optimised after removal of components that prohibit lyophilisation such as glycerol and betaine.
Equivalent Performance Between the LavaLAMP RNA Master Mix and RNA Component Kit
Figure 1. Comparing the performance of the LavaLAMP RNA Component Kit and the LavaLAMP RNA Master Mix. LAMP reactions (6 replicates per test) were set up using the indicated kits and targets (A, MS2 and B, Zika) with the same concentrations of Target-Specific Primer Mixes (1X) and the same target input amounts. The LavaLAMP RNA Component Kit reactions were formulated to 1X LavaLAMP RNA Buffer, 5 mM MgSO4 and 0.8 mM dNTPs (each) to match the final formulations of the LavaLAMP RNA Master Mix reactions. Green Fluorescent Dye was included in all reactions. Reactions were run on a CFX96 Thermal Cycler (Bio-Rad) at 68 °C for both the MS2 and Zika targets and fluorescence was measured during the course of the 60 minute reactions to determine TTR. NTC denotes No Target Control.
Faster, More Sensitive Detection with LavaLAMP RNA Component Kit
Figure 2. Performance comparison of the LavaLAMP RNA Component Kit vs. competitor kits. (A) RNA LAMP reactions were set up using the indicated kits according to manufacturer’s recommendations. Target MS2 RNA at varying input amounts, MS2 target RNA LAMP primers, and Green Fluorescent Dye (LavaLAMP Kit) were included in all reactions. Reactions were run on a CFX96 Thermal Cycler (Bio-Rad) at the following temperatures: LavaLAMP; 68 °C; other polymerases at the recommended 65 °C and fluorescence was measured over 60 minutes to determine the TTR. (B) Delta TTRs were calculated by subtracting the low target level TTR from the NTC TTR for each polymerase tested. Higher positive values indicate better signal to noise, and hence better performance. NTC denotes No Target Control.
Highly Consistent Assay Performance Between Operators
Figure 3. Analysis of operator to operator variability. On the same day, six replicate RNA LAMP reactions were set up per condition per by three different operators using the indicated amounts of MS2 target RNA, MS2 target RNA LAMP primers, and Green Fluorescent Dye. Reactions were run on a CFX96 Thermal Cycler (Bio-Rad) at 68 °C and amplification was monitored in real-time. Results were analysed to determine the TTR and averaged. NTC denotes No Target Control.
Consistent Day to Day Assay Performance
Figure 4. Analysis of day to day assay variability. On three different days, a single operator set up six replicate LAMP reactions per condition tested using the indicated amounts of MS2 target RNA, MS2 target RNA LAMP primers, and Green Fluorescent Dye. On each day the reactions were run on a CFX96 Thermal Cycler (Bio-Rad) at 68 °C, amplification was monitored in real-time. The results were analysed to determine the TTR, and the replicates were averaged and presented. NTC denotes No Target Control.
Licensing information: Lucigen is a fully licensed provider of LAMP reagents for research use. Patents WO 00/28082, WO 01/34790, and WO 01/77317 regarding the LAMP method are owned by the Eiken Chemical Co. Ltd. OmniAmp™ and Bst Polymerase, Exonuclease minus are sold under license for use in LAMP for research use only. The products may not be used for LAMP-based human or diagnostic purposes without obtaining a license from Eiken. US Patent 8093030 for OmniAmp DNA Polymerase is owned by LGC, Biosearch Technologies.
It is the sole responsibility of the buyer to ensure that use of the product does not infringe the patent rights of third parties. If the purchaser is not willing to accept these use limitations, LGC, Biosearch Technologies is willing to accept return of the product for a full refund.
SDS
- 30096 LavaLAMP RNA Component Kit
- 30097 LavaLAMP RNA Component Kit with Dye
- EU IT 30096 1 LavaLAMP RNA Component Kit
- EU IT 30097 1 LavaLAMP RNA Component Kit and Green Fluorescent Dye
MANUAL
- LavaLAMP RNA Component Kit
- LavaLAMP RNA Component Kit Quick Protocol
- LavaLAMP technical guide for new users
PIS