How do you reconstitute primers from IDT?
How do you reconstitute primers from IDT?
To obtain a 100 µM solution, multiply # nmol x 10. That will equal the # µL to use for resuspension. For example: 20 nmol X 10 = 200 µL. IDT offers a FREE online resuspension calculator and dilution calculator to make such calculations for you.
How do you dilute a TaqMan probe?
To achieve a concentration of 50 μM, a lyophilized primer with a mass of 119,000 pmol will be reconstituted with 2.38mL of 1X TE or sterile H2O. TaqMan® probes are commonly shipped in solution and their concentration is reported on the Applied Biosystems’ data analysis sheet.
How much water do I add to primer?
To determine the amount of water to add to the lyophilized primer simply multiply the number of nmol of primer in the tube by 10. That will be the amount of water to add to make a 100 µM primer stock. For example, if there are 38.2 nmol of primer a 100 µM primer stock is created by adding 382 µl of water.
How do you dilute oligos IDT?
Tips for resuspending and diluting your oligonucleotides
- During the dry-down process, oligos form a white flakey pellet at the bottom of the tube.
- If resuspension is difficult, try heating the oligo at 55°C for 1–5 minutes, then vortex thoroughly.
- IDT oligonucleotides (both DNA and RNA) are typically shipped dry.
How do you dilute oligonucleotides?
Alternatively, nuclease-free water, pH 7.0, can be used for resuspending oligonucleotides, but it will not modulate pH over time as will TE buffer. Use of HPLC- or molecular biology–grade water is preferable, as water from a deionizing system (such as Millipore) can be acidic, with a pH as low as 5.0.
How do you dilute a stock primer?
This can usually be found on the tube itself or the primer sheet supplied with the order. For every 1 nmoles, add 10 μL of PCR-grade water. For example, if a primer states 19.4 nmoles, then add 194 μL of PCR-grade water. Mix the solution by vortexing to reconstitute the primers.
How do you dilute the concentration of a primer?
How do you make a primer probe mix?
The units of a lyophilized primer or probe are given as a mass, in picomoles. To create a stock of primers or probe, one would reconstitute the primer or probe in sterile 1X TE (1mM Tris, 0.1mM EDTA, pH 8.0) or sterile, nuclease-free H2O.
How do I dilute my primer?
Can I dilute primer with water?
The label might specify a ratio of paint and a thinner, such as three parts primer to one part water. If so, pour the appropriate amount of primer and thinning substance into an empty bucket, instructs Acrylgiessen.com. Stir the mixture to blend it evenly.
How do you dilute a primer?
Can primers be Vortexed?
All Answers (1) Don’t vortex too much. But you can vortex gently for 5-10 for proper mixing. It would not break primers.
What are reconstituting and diluting primers and TaqMan® probes?
Reconstituting and Diluting Primers and TaqMan® Probes. Introduction. TaqMan® primers are commonly shipped in a lyophilized state. TaqMan® probes are sometimes shipped in the lyophilized state, but more often are shipped in solution (1X TE) and their concentration is reported on your Applied Biosystems’ oligofactory data analysis sheet.
What is optoptimized TaqMan®?
Optimized TaqMan® probe and PCR primer sets: TaqMan® Assays are formulated for use right out of the box. Life Technologies scientists have developed the most comprehensive set of real-time PCR assays available for gene expression, microRNA, noncoding RNA, protein analysis, mutation detection, copy number variation, and SNP genotyping.
How do I dilute the TaqMan® universal PCR master mix?
The TaqMan® Universal PCR Master Mix is at a 2X concentration and would therefore constitute half of the reaction or 25 μl in order to be at a 1X concentration. To dilute the primer and probe, use the following calculation. C1V1=C2V2
What is the mechanism of action of the TaqMan protein assay?
The mechanism of action is the same as that of the gene expression assay, using unlabeled primers and a TaqMan probe with a fluorescent label on the 5´ end and a nonfluorescent quencher (NFQ) on the 3´ end. Analyze TaqMan Protein Assay data using our free ProteinAssist™ Software package.