I am trying to isolate DNA from Halophyte using CTAB method, but the 260/280 ratio for the first time was below 1.8 and I have repeated the experiment. For this I have used the plant sample that kept in -80 °C but the nanodrop reading shows negative values and the ratio is above 2.
One month stored -80 degree can't influence DNA quality. You can't desire results due to not following properly extraction guidelines. It can happen for other reasons.
You may refer the following articles. Hope it may help you
https://www.ajol.info/index.php/ajb/article/viewFile/58477/46820
https://www.researchgate.net/publication/275330159_The_collection_and_storage_of_plant_material_for_DNA_extraction_The_Teabag_Method
https://www.researchgate.net/publication/267970290_A_simple_method_for_isolation_of_DNA_from_plants_suitable_for_long-term_storage_and_DNA_marker_analysis
@Yogesh Tiwari, Thank you so much are the articles are found to be helpful
I do not thing so. you can extract DNA by CTAB anytime that you want as long as your Samples store in -80.
@Mohsen Yoosefzadeh Najafabadi, Thank you. So the problem is associated with experimental errors? Attaching the gel image, anybody can suggest the possible problem.
Many Thanks
As Mr. Mohdmmad said one month stored -80 degree can't influence DNA quality. You can use DNA purification steps for qualification of your product.
There is no time limit for Plant part in -80. There could be problem with extraction method. Halophytes releases so much of phenolics that may interfere with DNA extraction. You improve your method so that it could be repetitive.
@ Khin Thuzar Win, Hukam Gehlot , Thanks for your reply. Why the bands become curved rather than a tight band?
Perhaps gel did not run properly. My be problem with electricity, fluctuations not vortex mixture/ Ladder properly. Such gel is called as smiling.
Hi
I saw this problem in extraction DNA from strawberry in due to high levels of phenols and arduous processes in extraction from freezing samples in contrast without any especial problem can extraction of DNA from fresh samples.
Well as far as I am know, the most important part in this process is the way that you extracting. probably your extraction was not quite good and it has some irrelevant material on it. you can extract again. However, this is an only suggestion.
Take care,
Mohsen
Thank You all for your valuable comments. I have repeated the procedure using silica dried material and the result was good
I have tested mine storage. year (-70), but quality or purity is important.
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