i need help with virus pcr - (May/30/2012 )
i'm new here and i have an problem:
i created baculogold viruses for infecting FS21 cells with a transfection kit from bd with recombinant dna. now i want to know if it is possible to make pcrs with this viruses. do the viruses crack during 95°C??
thanks for helping
It should be fine, most proteins will denature well at 95 deg C.
I am currently designing a protocol for HSV 2 Viral detection in mouse. I designed 2 different primers specific to HSV2 and mouse using NCBI primer-BLAST for SYBR set up without a probe (used for Taqman usually). HSV2 has dsDNA. I plan to run DNA directly on One step plus RT PCR with the primers and SYBR Green Master mix. Is this a correct and convenient protocol for my assay or do I need to create cDNA by first extracting RNA and then running them on Step One plus PCR?
I will also be working on another project to evaluate Clamydia levels in Beige mice. I want to be able to have similar protocols in terms of pre PCR processing. Kindly let me know how I can start working on this.
I selected SYBR as I have about 64