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(LAMP) Problems with Loop-mediated isothermal amplification

(LAMP) Problems with Loop-mediated isothermal amplification - DNA Techniques

(LAMP) Problems with Loop-mediated isothermal amplification - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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  #101  
Old 06-20-2013, 04:27 AM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Hi guys,
It is really helpful forum. I want to ask which dye are you using for real-time detection of LAMP and what is the recipe?
quick response will be appreciated.
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  #102  
Old 06-20-2013, 02:17 PM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

is someone using HNB? I'm trying to use 3 mM (1 microliter) but i didnt see change in color. does someone can send me the recipe ?? I will appreciate a lot. Also I was having goofd amplification results but now all my samples got positive. Do you have any idea how can I deal with this problem ?
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  #103  
Old 06-22-2013, 06:52 PM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Quote:
Originally Posted by MCaldera View Post
is someone using HNB? I'm trying to use 3 mM (1 microliter) but i didnt see change in color. does someone can send me the recipe ?? I will appreciate a lot. Also I was having goofd amplification results but now all my samples got positive. Do you have any idea how can I deal with this problem ?
False positive in your reaction are due to cross contamination of your LAMP product.

1. use different sets of pipettor & tips for prepare master mix and post LAMP reaction
2. After the reaction, Do not open the tubes at the same work place where you prepare your master mix
3.Replace all your reagents, decontaminate your pipettor and start fresh your reaction at new work station/ in biosafety cabinet.

Good Luck!
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  #104  
Old 08-16-2013, 10:53 PM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

For those using HNB to visualize your reactions, can you clearly see the colors at 120 uM HNB in 25 uL reaction? When I add 120 uM to my reaction solution, it looks completely clear. Not until 100 or 1000x concentration of HNB (12 or 120 mM HNB is 25 uL reaction) do I start to see colors that look similar to images in publications that use HNB. Any assistance would be very helpful.
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  #105  
Old 08-22-2013, 01:21 PM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Hi,
Can you help me? I'm using HNB with Cf: 120M, in the first I had good color and just 1h of amplification with F3, B3, FIP, BIP, so blue for + and purple for - ; and just one week later I haven't this change, I have already re prepare the stock of HNB but no reply for the color, however I always observed the bands whenever I migrate on gel;
Do you have some explain about that? is my stock of Hnb (I want mean one that is powder) is already inactive?
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  #106  
Old 09-10-2013, 08:26 AM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Hi guys,
I am finding limit of detection of LAMP assay.
LAMP was performed with serial dilution but I didn't get expected results. Endpoint of series was 10^-5 CFU/ml but at 10^-2 and 10^-3 had not products in both of color change and electrophoresis methods. (This series was checked with PCR on the same gene with LAMP and showed the expected results)
I did not know why it liked that.
So, can you help me?
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  #107  
Old 02-17-2014, 12:01 PM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Quote:
Originally Posted by Babak View Post
Anyone knows how to design primers for LAMP?

Thanks
Babak
Hi Babak,

Do you still face the same problem with primer design?

Thanks,

Ammar
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  #108  
Old 04-15-2014, 03:26 AM
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Default Re: (LAMP) Problems with Loop-mediated isothermal amplification

Quote:
Originally Posted by LAMP View Post
Hello there! I am also working with the LAMP primer design software. It opens and runs fine, but when I try to "Display" primers, nothing happens. Does anyone know how to solve this problem? Thank you!
Can you tell me the programme you use to design LAMP primer. Thanks a lot !
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amplificaion , amplification , dna , isothermal , lamp , loopmediated , primer design , problems


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