I would love to get some reassurance that these calculations are correct.
For example, I wish to make a 10X stock of the following:
20 μM of BIP/FIP,
10 μM of LF/LB,
2.5 μM B3/F3
I have 100uM stock for each primer. To a tube, I would add:
20ul FIP (100uM)
20ul BIP (100uM)
10ul LF (100uM)
10ul LB (100uM)
2.5ul F3 (100uM)
2.5ul B3 (100uM)
35ul Nuc Free Water/TE Buffer
Total : 100ul of 10X Primer Mix (20 μM of BIP/FIP, 10 μM of LF/LB, 2.5 μM B3/F3)
Thanks in advance :)
Your calculations for making a 10X LAMP primer mix seem correct based on the concentrations you provided. Here's the breakdown of the primer volumes you mentioned:
- 20 μM BIP: Add 20 μl of the 100 μM BIP stock.
- 20 μM FIP: Add 20 μl of the 100 μM FIP stock.
- 10 μM LF: Add 10 μl of the 100 μM LF stock.
- 10 μM LB: Add 10 μl of the 100 μM LB stock.
- 2.5 μM F3: Add 2.5 μl of the 100 μM F3 stock.
- 2.5 μM B3: Add 2.5 μl of the 100 μM B3 stock.
Finally, adjust the volume to 100 μl with nuclease-free water or TE buffer.
Overall, your calculation results in a 10X primer mix with the desired concentrations: 20 μM BIP/FIP, 10 μM LF/LB, and 2.5 μM B3/F3. This primer mix can be used for LAMP reactions at a 1X dilution. Remember to aliquot and store the 10X primer mix appropriately to avoid multiple freeze-thaw cycles, which may lead to degradation or loss of primer activity.
best..
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