Loop-Mediated Isothermal Amplification (LAMP): Primer Design and Assay Optimization

Nucleic acid based detection assays hold great promise for improved speed and sensitivity for a variety of applications such as microorganism detection, disease diagnosis and more. Loop-mediated isothermal amplification (LAMP), an isothermal nucleic acid amplification method which is rapidly gaining popularity, provides assay developers with a fast and cost-effective alternative to PCR for nucleic acid detection. LAMP’s isothermal characteristics allow for the development of simple, robust, low cost assays that can be deployed for point-of-care, point-of-need or environmental testing. Although a developed LAMP assay provides robust detection capabilities, initial design and development can prove difficult due to the 4 (or 6 for the faster assays) primer requirement. The need for 6 primers targeting 8 regions of the genome, the distance requirements between the primers, and sequence variability among clinical isolates of a given species put significant constraints on primer design. Improper primer design or selection leads to suboptimal assays resulting in slow amplification, non-specific amplification, and poor sensitivity. This webinar will review the basic mechanics of LAMP and how it may be used as a diagnostic assay. We will look in-depth at primer design with a focus on the use of primer design software and how specific settings improve primer design success specifically with LavaLAMP™ Enzyme-based Kits. We will also discuss optimization techniques to get the most out of your assay design.
This webinar will:
Review the mechanics of LAMP
Examine the primers required and their design
Learn how adjust software setting for better primer design
Demonstrate the effects of optimizing primer design on LAMP assay results
Explore additional ways to optimize LAMP assays
Check out our LAMP solutions here: www.lucigen.co...

[@katkat3458]

You explain very well! Thank you so much fo saving my thesis

[@testboga5991]

Thanks so much for the concise explanation! Best video on the subject I watched so far.

[@henrikslab]

Great video!

[@zaccomusic]

good contents friend

[@serhiysouchelnytskyi4697]

Thanks, informative webinar

[@MandeepSingh-lk2nz]

thank you for the informative content

[@mengistuhailemariam9050]

Many thanks

[@Soothyourminds]

Very informative, thank you

[@shalinipeeris9244]

Hi I am a bit confused at 60 -65 is DNA double stranded or single stranded, if it is double stranded how does FIP primer binds?

[@gutestederwelt]

at 60-65°C, the DNA is partially single-stranded (depending on its GC-content), which is enough for some primers to initiate the LAMP-reaction. I think the majority of the DNA remains double-stranded, but it doesn't matter. If your target sequence has a high GC-content, though, it may happen that 65°C is not enough for LAMP initiation and you would need to elevate temperature stepwise.

[@hailongwang1017]

Helpful, thanks

[@lubra9097]

Hey there, do you maybe have a good source for the tipical yields of PCR and LAMP, that you give in the video? Can't finde a citable one. Thanks in advance!

[@mengistuhailemariam9050]

could you come with Alethia LAMP? thanks

[@ciktiqshafaz]

Hi, i have question, why i got this "Could not generate Primer set(s). Please change parameters or target" statement when i tried to design my ITS region? I tried to do some changes in Detail Settings, but i still received that. Please help. Thanks in advance!

[@LucigenVideo]

Hello Atiquah! We would be happy to help with your primer design questions. Please contact us at [e-mail] as the root cause could be a number of things. Have a great day!

[@zachp2120]

No one voted in the poll 😢

[@krishnachandra3306]

how to design loop primer in detail explanation

[@henrikslab]

It should be a complimentary sequence to that region where you expect the loop to form