Want to calculate the percentage of transformation in plant, using agrobacterium mediated genetic transformation.
Usually two types of transformation frequencies are reported in Agrobacterium-mediated plant transformation. They are 1. Transient (GUS expression) transformation, 2. Stable transformation. These are calculated as follows
Mean frequency (%) of transient GUS expression = Number of explants producing blue spots/total number of explants X 100.
Mean frequency (%) of stable transformation = Number of transgenic plants regenerated/total number of explants X 100.
Hope this is helpful.
All the best!
1. Some people calculate it by:
Transformation efficiency (%) = [number of explants showing transformation/ number of explants inoculated] x (100%).
2. Some people calculate it by:
Transformation efficiency (%) = [Number of GUS positive plants / Number of calli inoculated with Agrobacterium] × (100%) [see attached paper, p.9]
3. Some people calculate it by:
"The transformation frequency was calculated as the number of kanamycin resistance plants/number of explants" [see attached paper, p.2 ]
Usually people calculate the transformation efficiency (%) as:
Transformation efficiency (%)= (Total number of PCR positive plants / Total number of inoculated callus) ×100.
Hope it may help you..
@ Yogesh,
One should be very careful in using PCR to identify 'positive' transgenic plants. You can very easily pick up the 'false positive' ones due to the exist of some resilient Agrobacterium (which is not eliminated during antibiotic selection). This can especially occur on the T0 putative transgenic lines.
Transformation efficiency in plant inoculated with Agrobacterium tumefaciens depend on number of Gus expression so the transformation efficiency can be calculated by divided the number of explant expressed with Gus on the total number of inoculated explant x1oo%
Hi everyone.
What is the method of determining the transformation efficiency of callus or regenerated plant (i.e. 50% that were not tested for GUS assay) ?
Usually two types of transformation frequencies are reported in Agrobacterium-mediated plant transformation. They are 1. Transient (GUS expression) transformation, 2. Stable transformation. These are calculated as follows
Mean frequency (%) of transient GUS expression = Number of explants producing blue spots/total number of explants X 100.
Mean frequency (%) of stable transformation = Number of transgenic plants regenerated/total number of explants X 100.
Hope this is helpful.
All the best!
How to calculate the transformation frequency/percentage in genetic transformation of plant?. Available from: https://www.researchgate.net/post/How_to_calculate_the_transformation_frequency_percentage_in_genetic_transformation_of_plant#view=593f996bcbd5c2d8c65e9502 [accessed Jun 13, 2017].
Normally, after plant transformation (e.g. callus); 50% of the infected calli undergo GUS assay, while the remaining 50% are co-cultivated / cultured for regeneration.
So, What is the method to determine the transformation efficiency percentage of the remaining 50% calli that were co-cultivated or transferred to regeneration media ?
Please, any body with knowledge on that method ?
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