In addition to IDT standard and custom mixed options for randomizing bases, we offer multiple base types (e.g., DNA, RNA, Affinity Plus™/locked nucleic acid, or 2'-O-methyl). This oligo has three insertions of a 50% G, 50% T mix (labeled K) and two insertions of a 10% A, 20% G, 70% T mix (labeled N1). This oligo has three insertions of a 25% for each base mix (labeled N) and three insertions of a 20% A, 20% C, 20% G, 40% T mix (labeled N1).Įxample 2: 5′ AGG (K:00005050)(K)(K) (N1:10002070)(N1)A GTA 3′ Each of these ratios must also include a unique name. Up to 4 unique custom-mixed ratios can be included in an oligo sequence. See below for what this looks like when incorporated in a sequence. Each mixed base in the sequence with this ratio can be notated with (N). Non-equimolar ratios will default to N1, N2, N3, or N4, based on the number of unique ratios in the sequence.įor example, an N mixed base with 25% of each base would be written as: (N:25252525). ![]() Instance of the custom mixed base must name and define the ratio, all subsequent identical insertions only need to include the name.įor equimolar ratios, our site will use only the IUB symbol to represent the ratio in the sequence. The ratio must be written in order of A, C, G, T (or A, C, G, U for RNA). If custom ratios are required, then for each mixed base position, the IUB code followed by a colon with a listing of the desired ratio are both included in parentheses. Custom mixed base services include additional fees in addition to the standard charges for synthesis and/or purification. Pricing for custom mixed bases is calculated by the number of unique ratios per order with a maximum of 4 distinct formulations per oligonucleotide. However, standard mixed bases will potentially have variation from the designated ratio due to the difference in coupling rates between base sites.īy requesting custom mixed bases, defined ratios can be added during synthesis, which includes unequal ratios. At the mixed base position(s), the synthesizer dispenses an equal ratio of the desired bases (e.g., a mix of A:C:G:T would be dispensed at 25:25:25:25, a mix of A:C:G would be 33:33:33:00, and a mix of A:C would be 50:50:00:00). Standard mixed bases can be inserted at any/all positions within a sequence at no additional charge. Standard mixing is a popular and economical way of incorporating mixed bases into oligo sequences. Two types of randomization are available-standard and custom mixed bases, both of which can be ordered through IDT’s online Oligo Entry tool by selecting the Bases tab. They can also be used to create diversity in clone libraries and in site directed mutagenesis. Mixed bases can be used for creating pools of oligos that include matches to a variable or unknown template sequence. The final order contains a population of oligos with a different base at the randomized position. IDT offers oligonucleotides containing randomized or "mixed" bases.
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