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PrimeTime Primer Probe Mix

IDT recommends that all probe melting temperature be approximately 6-8 degrees higher than primer melting temperatures. PrimeTime qPCR Probes do not contain Tm enhancing modifications such as Minor Groove Binders (MGB) that are commonly used in assays from Applied Biosystems. Therefore, directly entering of a probe sequence designed using Applied Biosystems software may have a reduced melting temperature within our assay, and performance may be significantly impacted.

Assay NameNotes

The Standard and XL qPCR Assays have options for primer to probe ratio and Dye-Quencher combinations. The actual amount of primer (in nanomoles) that will arrive in the tube is indicated below the sequence box for each primer.
# Bases: # Bases: Select the ratio of primer delivery amount to the probe delivery amount by moving the sliding scale.
5' 3'
Primer 1 delivery amount:
5 nanomoles
5' 3'
Primer 2 delivery amount:
5 nanomoles
Select a Ratio
PROBE   5' - 3' Dye-Quencher Mod View Table
# Bases: 

5' 3'
 Probe delivery amount:2.5 nanomoles

Multiple Entry
How many additional Assays would you like to create?
Upload your sequences

  1. Provide an Assay Name. This is the name that will appear on the product label. [Required]
  2. Enter any notes or comments about the assay. [Optional]
  3. Select a scale. [Required]
  4. Please use the following instructions to enter your sequences: [Required]
    • DNA bases: type the base in uppercase (e.g. A, C, T, G).
  5. Select the desired Dye-Quencher if applicable.
  6. Enter Mixed Bases. [Optional]
    • To enter a Standard Mixed Base, simply type in the respective IUB symbol. To obtain a list of these symbols, click the Mixed Bases tab above.
  7. Enter any notes or comments about the assay. [Optional]
  8. When you are finished entering the information, click the Add to Order button. You will then be taken to the Order Review Page.



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