Take 1 μl from the product of step 1 (saliva lysate) and add into the NASBA reaction mixture (without the enzyme mix) to make a total volume of 15 μl. Reaction mixture can either be prepared in-house or from the Life Sciences NASBA liquid kit (see Table 3 below) using one of the two temperature settings below.

*See table S1 for detailed composition.

1) Reaction mixture without the enzyme mix is incubated at 65°C for 2 min, followed by a 10-min incubation at 41°C. Following that, 5 μl of enzyme mix is added into the reaction and incubated at 41°C for a further of 90 to 120 min.

2) Alternatively, reaction mixture without the enzyme mix is incubated at 95°C for 5 min, followed by a 10-min incubation at 41°C. Following incubation, 5 μl of enzyme mix is added into the reaction and incubated at 41°C for a further 90 to 120 min.

A fluorescence plate reader (e.g., FLUOstar) can be used to monitor the reaction in real-time or as an end point assay.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.