Hello Everyone
1. When we use DSS Salt for Colitis Induction then there is degradation of RNA (Its documented in various articles) content in Intestine. so it will affect your RT-qPCR results as low Ct values which will mislead you. someone will say you can compare with Experimental control group. but that is not a very reliable way, if someone know any KIT or any method to keep RNA Integrated and so so no issue with RT-qPCR then please suggest. . How to avoid or overcome RNA degradation in DSS model?
2. DSS either have an effect on Protein quality or not?
Simon Keely
Thank You.Had overcome the issue up to some extent and had followed to Transcriptom as well; and it worked well. We adopted the following ways.
1. Used a different RNA extraction Kit "TransZolUP Plus RNA Kit", Catalog Number：ER501-01" https://www.transgenbiotech.com/rna_purification/transzol_up_plus_rna_kit.html.
2. Washing tissues several times to clean up DSS.
Now we got CT values around 14-25 in RT-qPCR and seems reliable now. We are trying to improve as this time we are using the cDNA we make; and we do not make dilution; otherwise we may not be able get reliable CT values. This is time consuming and not economical to make cDNA again and again; as sometime we need to check many genes.
I would like to know detail method that you applies in lithium chloride clean up. Pease share any detail method. Thanks. email: [email protected]
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