It´s a great effort this discussion, but i think it´s necessary more time for every module
and include all the information, or maybe a particular discussion for only one module.
At this time, much countries have a really good level for LMO detection, and we use this information to go the next step.
We add specific comments and suggestions directly in the text.
posted on 2018-04-13 18:50 UTC by M. en S. MARIA GUADALUPE BARRERA ANDRADE, Mexico
Dear M. en S. Maria Guadalupe Barrera Andrade of Mexico, thank you for adding information about leaves. This matrix has not been sufficiently well considered in the manual.
I have a few additional suggestions regarding sampling handling (text added around line 135). For protein-based testing it is not necessary to grind the leaves in liquid nitrogen, nor to freeze them. They can be sent on ice to a testing location, or better still, tested in the field. The samples can be extracted directly by adding sample buffer.
It is not normally feasible to freeze samples in liquid nitrogen and grinding in high throughput commercial laboratories – this approach can lead to low throughput of samples. Grinding of seed and grain samples at room temperature is usually sufficient, and small sample can be taken and re-ground if it is desired to extract very small samples such as 100mg of powder when looking for low levels of LMOs.
I do not understand the basis of the minimum number of leaf pieces – the sample could be for example a leaf from a plant identified as a LMO and that the PCR is being used solely for identification of the specific LMO, in which case multiple leaf pieces would not be needed.
In addition for protein-based methods, because they can be performed relatively inexpensively, it is not usual to bulk 100 leaf pieces (table 3) – although if a method validation has been done, such an approach can be used; the manufacturers of the kits do not include this approach in their instructions. It is important to make sure the readers of the document do not confuse the processes used for PCR methods with those used for protein-based methods.
We also support the inclusion of a section on Verification of DNA integrity. I am not sure whether this is dealt with elsewhere in the modules, and there is not time now to check.
posted on 2018-04-15 20:50 UTC by Dr Raymond Shillito, Bayer CropScience
Thank you very much for your input. We acknowledge the challenge in reviewing the manual in its entirety in the time provided, unfortunately however, it is not possible to provide more time for the discussions.
We thank you for your understanding.
(edited on 2018-04-16 01:14 UTC by Dina Abdelhakim)
posted on 2018-04-16 01:08 UTC by Dina Abdelhakim, SCBD