How do you order primers?
How to design and order Primers
- Length:
- Primers with long runs of a single base should generally be avoided.
- Primers should have a GC-content between 50 and 70 %
- If possible, primers should be stickier at the 5′ ends than at the 3’end.
- Primers should not contain complimentary sequences (palindromes) within themselves.
How do you order oligonucleotides?
Log into your IDT web account using your login ID and password. From the “Order” tab select “Main Menu” and choose the product type you need. For custom oligos, you will ned to provide a sequence name for the sequence entered and a synthesis scale and purification type.
How do you reconstitute primers Invitrogen?
For example, if there are 38.2 nmol of primer a 100 µM primer stock is created by adding 382 µl of water. The original primer tubes are used for this 100 µM stock. Master stock primers newly suspended in water should be allowed to sit at room temperature for 10 minutes before they are used for working stock dilutions.
How do you make a primer for PCR?
PCR Primer Design Tips
- Aim for the GC content to be between 40 and 60% with the 3′ of a primer ending in G or C to promote binding.
- A good length for PCR primers is generally around 18-30 bases.
- Try to make the melting temperature (Tm) of the primers between 65°C and 75°C, and within 5°C of each other.
How do you design forward primer?
Forward and reverse primers should be about 500 bp apart. The 3′ end of the primer should be a G or a C. The genomic sequence that comes from the computer is just one strand; the complementary strand is not shown. For the forward primer, you can use the sequence directly.
What is oligonucleotide primer?
The term oligonucleotide is derived from the Greek “oligo,” which means few or small. Oligonucleotides made up of 2′-deoxyribonucleotides are the molecules used in polymerase chain reaction (PCR). These are referred to as primers and are used to massively amplify a small amount of DNA.
How much does a PCR primer cost?
Service Details
| Service Name | Unit | Unit Price |
|---|---|---|
| Primer Synthesis (25 nmol) | 1 Base | $0.53* |
| Primer Synthesis (100 nmol) | 1 Base | $1.05* |
| Primer Synthesis (250 nmol) | 1 Base | $1.88* |
| sgRNA PCR Primer Pair Design & Synthesis Service (25 nmol/primer) | 1 Pair | $148.50 |
How do you resuspend dry oligos?
Oligo resuspension
- 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.