Hello,
It is the first time I post question in here. For my master thesis, I am studying the effect of medium composition in the number of embryos that germinates after rescuing them. I used 5 cross combinations and pollinated 20 flowers for each cross combination and repeated that 3 three times but unfortunately one of my repetition can't be used (technical problems). I rescued the embryos from these repetitions separately, so can it be considered as a technical replicate? Most of the publications only says that they consider 12 replicates but i am not sure what they mean. Indeed, in addition of this two repetitions, I have around 15 embryos per media tested which would be 30 embryos in total per media with the two repetitions. So is it possible that considering each embryo seperatly I can perform statistical analysis ?
In addition, 6 pollinated flowers were harvest 8 DAP, 6 others 12 DAP and the last 8 were used for the seed set. So everytime I performed embryo rescue, I bulked the embryos from the six (or less if the cross wasn't a success) flowers that were harvested, is that my biological replicates ?
I am a little bit confused with all of this.
Hi Diana,
Well, at first the difference between biological and technical replicates: A biological replicate is an independent sample or observation. On the contrary a technical replicate is a repetition of the same sample used in the same way, in theory if you perform experiments with technical replicates you should obtain the same result because you are using the same sample and same conditions. Technical replicates are done to minimize human error while doing experiments.
This is very important in quantitative experiments. For example a sample of DNA is a biological replicate. If you use the sample to repeat three times the very same experiment you have three technical replicates. If your experiment involves measuring you can now calculate the average between the technical replicates.
In statistics as Ette said, the more samples (biological replicates) you have, the more your results will be likely to be true, because it is less probable that you found that result just by chance.
So, i think that each of your flowers and embyos are biological replicates (usually i say replicates for biological replicates and repetitions for technical replicates). From your biological replicates you can do how many technical replicates you want, your result will be more precise (even if you are not doing a quantitative experiment, e.g. you will obtain more positives).
If you consider 15 embryos a group of biological replicates and so you have 15 samples in two experiments yeah, that is fine. As Ette said, no standard number exist. On the contrary, the more the better.
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