We have a project on development of a dry loop mediated isothermal amplification (LAMP)-based kit for detection of plant viruses on field site. One of the challenges in this project is the lyophilization of LAMP mix in 0.2 mL PCR tubes. For reference, I am using a FreeZone 2.5 Liter benchtop freeze dryer. But any suggestion even if you're using a different brand/model will be much appreciated.
I think the brand or model of the freeze dryer does not have an effect on the sample to be processed. It is the recommended physical conditions and built-in performance of the freeze dryer that count in order for you to come up with the processed samples for the dry LAMP.
Can you go over the volume of the sample for freeze drying. Don't you think it is too small for processing.
Hi Cris
Can you help me here with more information on what exactly is the challenge you are facing in lyophilization!?
What we do here at my lab is, we freeze the sample to -80 C before processing the sample in a freeze dryer at -60 C. This step is very crucial, when you keep the sample in lyophilizer it should be in solid-state ( ice form) so that the sublimation process can take place ( solid to gas ). So my suggestion is, ensure that you are samples are frozen before placing inside the lyophilizer, and take precaution while using a 0.2 ml tube, since the volume will be less, there are high chances of it getting melted.
Good luck!
Dr. Gemerlyn Garcia Yes, I just mentioned the brand/model just in case someone is using the same brand as mine who can provide the most relevant suggestion. But suggestion from users using other brands is also equally important. Regarding the sample volume, we do wish to use just 10-uL in order to diminish the cost of the assay when it is already adapted in the field. But you are right, upon reading your comment and Mr. Kumar's answers, it seems utilizing that small volume will become problematic during freeze drying as it will hinder the sublimation process. I guess if I really want to lyophilize 10uL volumes, I should look for ways to prevent the samples from melting and therefore enhance sublimation.
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