The forward primer was provided by the manufacturer as a lyophilized powder. If the manufacturer delivers 127.4 nmoles of this primer, what volume of buffer should you dissolve it in to make a 100 μM stock?
Assuming the buffer has a density of 1 g/mL, the volume of buffer needed to dissolve 127.4 nmoles of the primer in order to make a 100 μM stock would be 127.4 μL.
To determine the volume of buffer needed to make a 100 μM stock solution from a given amount of lyophilized primer, you can follow these steps:
Step 1: Determine the molecular weight of the primer.
If the molecular weight of the primer is not provided, you can calculate it by adding up the atomic masses of each element in the primer's chemical formula.
Step 2: Convert the amount of primer from nanomoles (nmoles) to moles (μmoles).
To convert nmoles to μmoles, divide the given amount of primer by 1000 (1 μmol = 1000 nmol).
Step 3: Convert the stock concentration from μM to M.
To convert from micromolar (μM) to molar (M), divide the desired concentration by 1,000,000 (1 M = 1,000,000 μM).
Step 4: Calculate the total amount of moles needed for the stock solution.
Multiply the converted amount of primer in moles (from Step 2) by the desired stock concentration in moles (from Step 3).
Step 5: Calculate the volume of buffer needed.
Divide the total amount of moles needed (from Step 4) by the desired stock concentration in M (from Step 3). This will give you the volume of buffer required in liters.
Let's assume the molecular weight of the primer is 1000 g/mol, and the desired stock concentration is 100 μM:
Step 1: Molecular weight = 1000 g/mol
Step 2: Conversion from nmol to μmol
127.4 nmol = 127.4/1000 μmol = 0.1274 μmol
Step 3: Conversion from μM to M
100 μM = 100/1,000,000 M = 0.0001 M
Step 4: Total amount of moles needed
0.1274 μmol x 0.0001 M = 0.00001274 mol
Step 5: Volume of buffer needed
0.00001274 mol / 0.0001 M = 0.1274 L = 127.4 μL
Therefore, you would need to dissolve the lyophilized primer in 127.4 μL of buffer to make a 100 μM stock solution.
To determine the volume of buffer needed to dissolve the lyophilized forward primer and make a 100 μM stock, we will use the following formula:
Stock Concentration (M) = (Amount of Primer (moles)) / (Volume of Buffer (Liters))
First, we need to convert the given number of nanomoles (nmoles) to moles:
Given: Amount of Primer = 127.4 nmoles
Conversion: 1 mole = 10^9 nmoles
Amount of Primer (moles) = 127.4 nmoles / (10^9 nmoles/mole)
≈ 1.274 x 10^-7 moles
Next, we can insert this value along with the desired stock concentration into the formula and solve for the volume of buffer:
100 μM = (1.274 x 10^-7 moles) / (Volume of Buffer)
Rearranging the equation to solve for the Volume of Buffer:
Volume of Buffer = (1.274 x 10^-7 moles) / (100 μM)
Since 1 μM = 10^-6 moles/Liter, we can convert the stock concentration:
Volume of Buffer = (1.274 x 10^-7 moles) / (100 x 10^-6 moles/Liter)
= (1.274 x 10^-7) / (10^-4)
= 1.274 x 10^-7 / 10^-4
= 1.274 x 10^-7 x 10^4
= 1.274 x 10^-3
The volume of buffer required to make a 100 μM stock is approximately 1.274 μL.