No discussions found
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Found 3 matching solutions for this experiment
Upstream tips |
Protocol tips |
Downstream tips |
|
For RNA to dissolve better and to yield high levels, preheat elution buffer at 95C.
Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation.
To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
Do vortex for 2-plus minutes. This will facilitate the removal of tightly bound proteins typically associated with RNA. |
|
Protocol tips |
For RNA to dissolve better and to yield high levels, preheat elution buffer at 95C.
Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation.
To capture a higher amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step.
Do vortex for 2-plus minutes. This will facilitate the removal of tightly bound proteins typically associated with RNA. |
Upstream tips |
Protocol tips |
Downstream tips |
|
Total RNA was isolated from frozen tissue using the Promega Total RNA Isolation System (Promega, Madison, WI, USA). For cDNA synthesis, 4–8 μg of total RNA was reverse transcribed with Superscript III reverse transcriptase using an oligo-(dT)12–18 primer according to the supplier’s protocol (Invitrogen, Carlsbad, CA, USA). A single cDNA preparation from each specimen was used for the assay of all antimicrobial products tested. |
|
Protocol tips |
Total RNA was isolated from frozen tissue using the Promega Total RNA Isolation System (Promega, Madison, WI, USA). For cDNA synthesis, 4–8 μg of total RNA was reverse transcribed with Superscript III reverse transcriptase using an oligo-(dT)12–18 primer according to the supplier’s protocol (Invitrogen, Carlsbad, CA, USA). A single cDNA preparation from each specimen was used for the assay of all antimicrobial products tested. |
Upstream tips |
Protocol tips |
Downstream tips |
Be careful to create an RNase-free working environment |
Always mix the sample tube well after addition of each reagent. |
|
Upstream tips |
Be careful to create an RNase-free working environment |
Protocol tips |
Always mix the sample tube well after addition of each reagent. |
Can't find the product you've used to perform this experiment? It would be great if you can help us by
Adding a product!