Combining RNA from technical replicates of one biological sample is a common and valid approach for 5' Rapid Amplification of cDNA Ends (RACE) experiments. This practice can enhance the robustness and reliability of the experiment by minimizing variability introduced during sample preparation and processing. Key considerations include ensuring consistency in RNA preparation methods, normalizing RNA concentrations if needed, verifying RNA quality, designing experiments to account for technical variability, and interpreting results appropriately, considering potential technical artifacts. Overall, combining RNA from technical replicates can improve the reliability of 5' RACE experiments, provided that proper experimental design and data analysis techniques are employed.

Hello,

It should be fine to combine samples for RACE. In case the gene undergoes splicing towards 5'-end, you may to be careful sometimes.

Hi, joining to upper comments: when you goes to combine sample you can improve 5'RACE, but if your target transcript low in many samples, without normalization of cDNA - you can decrease output from reaction. And after 5'RACE through combined samples you more often need cloning (and handle more clones to work) output product to take sequence.

for me it is rarely worth to pool samples: when you mix a good sample with a bad sample then you get .... a bigger bad sample. So exept if you do not have enough RNA with one sample to do the experiment then do not pool