Just to make it easy for you-check the nm on the vial of the primer. Suppose to find 27.4 nm or so. Multiply by 10 and will be 274 uL. This is 100uM Stock. In an eppendorf tube add 80 uL H2O and then 10 uL from the diluted forward and backward primers then you will have 10uL working solution. Do not forget to mixed and spin each time. Good luck!
How to make 100um stock of primers?
Just to make it easy for you-check the nm on the vial of the primer. Suppose to find 27.4 nm or so. Multiply by 10 and will be 274 uL. This is 100uM Stock. In an eppendorf tube add 80 uL H2O and then 10 uL from the diluted forward and backward primers then you will have 10uL working solution.
How to make 100 um stock solution?
To make the 100 uM stock, multiply this by 10 and add than many ul of water (e.g. 233 µl for 46.6 nmol = 200 µM stock solution). 6. Write the name of the oligo on the cap of the stock tube. Then put the original stock in a common oligo box (normally large fluorescent boxes in tall freezers in Trude's or Pat's drawer). 7.
How to prepare 1nanomole of primer for 100 pmol main stock?
For 100 pmol main stock you can add 255 µl either sterile mill-Q water or 10 mM TE (pH-8). Spin the primer tubes at 10k for 15 mins. Dilute to 1nanomole concentration.
How do I Add H2O to my primer stock?
The pellet can often come dislodged during shipping and may be in the cap! To determine the amount of H2O to add to the lyophilized primer simply multiply the number of nmol of primer in the tube by 10 and that will be the amount of H2O to add to make a 100 µM primer stock.
How do you make a stock primer?
PrimersAdd sterile ddH 2 O (using barrier tips) to resuspend the dried primer to a concentration of 100 μ M. ... To make a 10μM working solution of primer, dilute the 100μM stock 1:10 with sterile ddH 2 O. ... 10μM primer solution is diluted 1:25 in a PCR reaction.
How would you prepare 500ul of a working stock of 100 nm primers from a stock of 10?
If you have 100 umoles in 500 uL, that is 200 nmoles per uL. If your reaction needs to contain 500 nmoles, you will need 2.5 uL of your stock solution per reaction. If however your stock solution is 100uM (micromolAR), then you will need to dilute it 1/5000 to achieve 500nM (nanomolar).
How much water should you add to 10 Umole primer to make a 100 mM solution?
For example, for an amount of 20.91 nmole of lyophilized primer (the amount is provided by the vendor), I add 209.1 ul water or TE buffer and get 100 uM stock solution. To prepare 10 uM working solution, I take 10 ul stock solution (at 100uM) and add to 90 ul water.
How is 100uM calculated?
Making a 100 μM solution To resuspend your oligonucleotides to 100 μM, simply multiply the number of nanomoles (nmol) by 10 to get the volume (in μL) of water or buffer to add. The equation on the left shows an example, assuming you have an oligonucleotide of 1 nmol final yield.
How do you make 100 microMolar solutions?
For a 1:100 dilution, add 1 mL of the stock solution to 99 mL solvent. (ii) The second dilution. Add 1 mL of the 10 mmol/L solution to 99 mL solvent.
How do you make a 100 mM stock solution?
For practical purpuse it is best to prepare a concentrated stock solution. 416.6 mg in 10 ml will give you 100 mM solution. This can be aliquoted into multiple vials and kept frozen. When needed, thaw one vial and then use this stock soluition to make further dilutions.
Can I dilute primer with water?
you can use distilled water for primer dilution. The exact answer is given by Paul! 1XTE is the solution for your problem for 100%.
How do you dilute a stock primer solution?
To make a typical 100 microMolar (100X) stock concentration of primers, dissolve the primers in a volume of sterile distilled water that is 10X the amount of nmoles in the tube, using microliters of water. This value is printed on the side of the tube.
How do you do a 1 100 dilution?
For a 1:100 dilution, one part of the solution is mixed with 99 parts new solvent. Mixing 100 µL of a stock solution with 900 µL of water makes a 1:10 dilution. The final volume of the diluted sample is 1000 µL (1 mL), and the concentration is 1/10 that of the original solution.
What is this μm?
micrometre, also called micron, metric unit of measure for length equal to 0.001 mm, or about 0.000039 inch. Its symbol is μm. The micrometre is commonly employed to measure the thickness or diameter of microscopic objects, such as microorganisms and colloidal particles.
How do I make a 10 mg/ml stock solution?
To prepare a concentration of 10 µg/ml, pipette out 10 µl of the drug in a test tube using a micropipette. Then dilute it with 990 µl (making a total volume of 1 ml) of the solvent (ethanol, methanol, water etc) you are going to use in your experiment.
How much primer do I need for PCR?
for 10 pico mole concentration of primer for standard PCR from that stock you have to take 1 microlitter of each primer for 100 micro litter reaction.
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After adding the TE buffer to the primer, where will the primer be stored?
Popular Answers (1)
To make a typical 100 microMolar (100X) stock concentration of primers, dissolve the primers in a volume of sterile distilled water that is 10X the amount of nmoles in the tube, using microliters of water. This value is printed on the side of the tube.
All Answers (28)
To make a typical 100 microMolar (100X) stock concentration of primers, dissolve the primers in a volume of sterile distilled water that is 10X the amount of nmoles in the tube, using microliters of water. This value is printed on the side of the tube.
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Everyone has explained very well. Additionally, I think following links are very helpful in calculating accurately.
Similar questions and discussions
How do I get 200ng of oligonucleotide from 100uM oligonucleotide stock?
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Before you start
So you have designed and ordered your primers and now they have finally arrived. But how do you prepare lyophilised (freeze-dried powder form) oligo primers for use in PCR? If you are unable to prepare the primers straight away, then store them at 4oC until the time comes.
1. Reconstitute your stock primers
First things first, you should briefly (approximately 30 seconds) centrifuge your primers to pull all of the lyophilised powder to the bottom of the tube. Otherwise, if the powder is stuck in the lid while opening the tube, you could potentially damage your primers.
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Most PCR reactions use 0.1−0.5 μM primer. Assuming a maximum concentration of 0.5 μM and a reaction volume of 20 μL, each reaction will require 10 pmol oligonucleotide primer. Bhoomika Sharma
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If you don't set the volume of PCR reaction mix, one can't answer the question in term of volume! Dilution factor will be 50 starting from your 10µM solution.
