Human TCR primers and PCR protocol


#1

Does anyone have experience with the BIOMED2 TCRB primers? Are these considered the best ones to use? Right now I just want to implement a feasibility study on new sample types to determine whether there is enough rearranged TCRB DNA present in the sample to make it worth it to spend the money for repertoire sequencing. So we will probably just do PCR and then run the product out on a gel to determine if we get right sized bands.

Any suggestions welcome. Thanks!


#2

We have done some single-cell work based on the primers by Han et al., which are ok. As long as your input material is gDNA and don’t care about allelic variation in TRBV, there is probably nothing arguing against BIOMED-2 (as this is what they were designed for). However, if you want to have (nearly) full-length V segment information, Han et al. is a good starting point. Note that the diversity between segments is higher for TCR than for Ig, so it is rather challenging to find conserved sites, i.e. you will often end up with large primer panels. If you are working with cDNA using a constant region primer will save you the hazzle of dealing with different J segments (especially TRAJ).


#3

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