PEG6000沉淀法浓缩的病毒测滴度,0.1 μl和2 μl组阳性率只差了2倍
求助各位大侠了,我用PEG6000沉淀法浓缩病毒,操作如下(按照别的实验室的protocol来的):
9 vol medium
2 vol 50% PEG6000
1 vol 4M NaCl 混匀以后,4℃静置过夜
1. Spin the 50ml tube at 3000rpm, 4 degree for 30min,
2. Carefully dump most of the supernatant (should see white pellet on the bottom, PEG precipitate with virus)
3. Spin again at 3000rpm, 4 degree for 5min,
4. Remove the rest of supernatant
5. Use 600ul PBS to resuspend the pellet, pipetting up and down to gently mix well (avoid too many bubbles), leave in 4 degree for at least 2hr
我最后是用DMEM溶解的,大概浓缩了70倍。在NIH-3T3里检测滴度,平行做了3组,操作应该是没问题的。滴度检测结果如下(病毒感染NIH-3T3细胞4h后换液):
0.1 μl病毒阳性率:5% 结合浓缩倍数,这一组的效价跟我浓缩前是接近的。
0.25 μl病毒阳性率:5%
1 μl病毒阳性率:8.5%
2 μl病毒阳性率:10%
问题:不同梯度,效价差别不大,为什么呢?(浓缩70倍合适吗?PBS比DMEM更为合适?)
求助各位大侠啦,在下感激不尽!
最后编辑于 2022-10-09 · 浏览 7276