Hi Ruchita. For gene expression, it would also work well if you could design primers by urself. Just open nucleotide sequence in NCBI and click pick primers option. It will give you 5-10 pairs of primers. Pick the best ones using product size, self complemenrarity and 5’ complimentarity as important criteria.

good luck

Hi Ruchita,

It does not matter Wistar rat or Lewis or Fisher rat, primer sequences you will design as Ravi said will work for Wistar rat. Try to test 2 pairs for each gene, choose the one which has best melting curve. Did you assess osteochondral defect by histology also if the defects were filled with cartilage or fibrous tissue after stem cells therapy.

Best,

Subhash

In general as you are using stem cells it might be worth looking at klf4, runx3 also add some bone markers such as runx2,sp1, alp to ensure you know what lineage your stem cells are differentiating.

@ Ravi Kumar Sharma: Thank you so much for your reply. Initially, I thought of trying the sequences already in literature and then trying with its designing.

@ Subhash Juneja: Sir, thank you so much for your reply! I have still not assessed the repair of osteochondral model with stem cells. But, its under the pipeline. We have successfully developed the model. Now, we are waiting for the efficacy studies of stem cells to be done. That is the reason, I wanted some supporting data along with histology results to prove the cartilage regeneration.