I am working on T1 transgenic rice plants. Let's say that for one of the transgenic lines, I have only one positive plant (pcr tested to contain the inserted gene). I need to take leaf samples for further test to quantify cellulose and cellulase and do q-pcr. Since I only have one plant, what should I do about replication? If I take 3 leaf samples from that plant, is it considered as biological or technical replicate?
For T1 transgenic plants, since there might be high variation between different leaves even within the same plant, is it ok if I pick a few leaves, freeze in liquid nitrogen, grind into powder and mix them up, then take 3 portions out as replicate for q-pcr analysis?
Fr om the same single transgenic plant, taking different leaves could mean same sample. If you get the seeds, seeds could be different samples.
Hi,
a technical replicate is used when is necessary to test the same sample multiple times. Generally you might think of a technical replicate when you are testing the variability in the testing protocol.
A biological replicate, instead, is used when performed the same test multiple times on the same material (cells, tissue etc.). Biological replicates can be used to test the variability between samples selected on the basis of being otherwise identical.
If you would like to test the same prep multiple times (from 500 uL of DNA you aliquote to 5 x 100uL each), than it is considered as a technical replicate.
If you instead are taking multiple samples and prepare each of them following the same protocol, then you have a biological replicate.
Technical replicates are helpful to quantify the internal variation of your test, while the biological replicates enables you to observe the variation between samples.
In your specific case, 3 samples are considered as biological replicates.
Best regards
If it would not be rize...
I would recommand to take 3 leaves:
The basical part of each leaf you put on replication medium in order to get new plantlets. The upper part you can evaluate in PCR. Label them 1 and 2 and 3
- Badges
- Science topic
- Similar topics
- Methodology
- Research Methodology
- Research Papers