Q
For Stereo-seq Transcriptomics experiments with fresh frozen samples, is it recommended to do multiple replicates?
A
It is recommended to determine whether it needs to be repeated according to the purpose of the experiment.
Q
How long does the Stereo-seq experiment take?
A
The workflow of Stereo-seq Permeabilization Set takes around 4h;
The workflow of Stereo-seq Transcriptomics Set takes around 9h;
The workflow of STOmics Library Preparation kit takes around 5h.
Q
What is the shelf life of the reagents?
A
Please refer to the kit label.
Q
Which type of samples are currently suitable for Stereo-seq Transcriptomics Set?
A
The set is applicable to all common animal samples.
Q
What should be paid attention to during tissue embedding?
A
The tissue size should be no more than 0.9 cm x 0.9 cm x 2cm, and appropriate cryomold or stainless- steel based mold should be chosen based on the tissue size. To avoid RNA degradation, tissue embedding should be finished within 30 min after tissue harvest. Excess liquid on the tissue should be removed to avoid ice formation during embedding. Air bubbles should be avoided when filling the OCT in the Cryomold or stainless-steel.
Q
What is the cause for serious distortion of the tissue during embedding?
A
The distortion of tissue could be caused by tissue compression during tissue harvest. In this case, tissue harvest method should be adjusted accordingly. The volume expansion of the OCT during freezing could also lead to tissue distortion. The embedding conditions should be adjusted according to the specific situation, such as, considering changing Cryomold or stainless-steel based mold size or material.
Q
How to avoid ice crystal formation within the tissue?
A
Excess liquid on the tissue should be removed before freezing. Rapid freezing is recommended to avoid ice crystals in the tissue since slow freezing produces large ice crystals.
Q
Will the amount of OCT affect the tissue during embedding?
A
Tissue may be squeezed when submerged in excess OCT, especially for small sized tissue. Therefore, it is recommended to choose an appropriately sized Cryomold or stainless-steel based mold
.
Q
Does OCT affect RNA extraction and RIN evaluation?
A
Yes, excessive OCT will affect the result of RNA extraction.
Q
How to avoid mRNA degradation during fresh tissue preparation?
A
It is strongly recommended to perform tissue embedding within 30 minutes after tissue harvest to avoid tissue RNA degradation to the greatest extent.
