Hi there,
I want to ask whether a ratio of 260/230 of 1,55 is considered low for primer after its resuspension.
Thanks in advance
It depends on what you dissolve the primer in and the primer sequence. Salts absorb at 230 and will lower the 260/230 ratio but if we assume that you dissolve in water or TE then this will not be the case. I think that the oligo sequence is the likely answer. The 260 measurement in oligos depends on the nearest neighboar bases as well as the actual base so the sequences cc , ct, tc and tt have low extinction coefficients while the sequences aa or ag or ga have high extinctions. It may just be that your oligo is rich in As and Gs giving a high 260 overall value which would mean a low 260/230 ratio
generally a ratio between 2.0-2.5 is considered good indicating absence of unwanted compound impurities. But as mentioned by paul it basically mainly depends on primer solvent and its sequence.
To determine whether a ratio of 260/230 of 1.55 is considered low for a primer after resuspension, we need to understand what the 260/230 ratio represents and what is considered an acceptable range.
The 260/230 ratio is a measurement commonly used in molecular biology to assess the purity of nucleic acids, such as DNA or RNA, after purification or resuspension. Specifically, it compares the absorbance at 260 nm (which corresponds to nucleic acid concentration) to the absorbance at 230 nm (which indicates the presence of contaminants like proteins or organic compounds).
In general, a high 260/230 ratio indicates that the sample is relatively pure, while a low ratio suggests the presence of contaminants. However, the acceptable range for the 260/230 ratio can vary depending on the specific application and the guidelines provided by the manufacturer of the primer or the purification kit.
For most molecular biology applications, a 260/230 ratio above 1.8 is typically considered good and indicates a relatively clean sample. Ratios between 1.5 and 1.8 are usually acceptable, but they may indicate the presence of some contaminants. Ratios below 1.5 are generally considered low and may suggest the presence of significant contamination, which could affect downstream applications.
Based on this information, a 260/230 ratio of 1.55 would be considered relatively low. It suggests that there might be some contaminants present in the resuspended primer sample. If possible, it is recommended to re-purify or clean up the primer to improve its quality before using it in downstream applications.
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