不同电击管的电压参数需要换算,时间相同。以20ul电击管参数为基准,100ul比20ul约上调10V;200ul是20ul的约2.4倍;1000ul是20ul约2.5倍;120ul的是20ul的约1.4倍,600ul的是20ul的约1.6倍。下面是转换示例:
20ul:500V
100ul: 510V (20ul电压+10V)
200ul:1200V (20ul电压的2.4倍)
1000ul:1250V (20ul电压的2.5倍)
120ul:700V (20ul电压的1.4倍)
600ul:800V (20ul电压的1.6倍)
*部分单核细胞相关的细胞系,电转质粒后几乎所有电转成功的细胞会死亡,而电转mRNA,siRNA和Cas9 RNP没关系。Celetrix公司专门开发了试剂盒可以帮助单核细胞类的细胞系进行质粒电转,目前验证过THP-1和U937这两种细胞系电转效果非常好,而Molm-13和Sup-M2细胞电转质粒细胞死亡问题还没有解决。
Cell Type | Mode | 20ul, 100ul | 200ul, 1ml |
---|---|---|---|
Human PBMC | PBMC | 1200 -1400 | NA |
Human T cells, unstimulated | PBMC | 1300 -1450 | NA |
Human B cells, unstimulated | PBMC | 1300 -1450 | NA |
Human NK cells, unstimulated | PBMC | 1300 -1450 | NA |
Human Neutrophil | PBMC | 1300 -1450 | NA |
Mouse lymph node primary T cell | PBMC | 1300 -1450 | NA |
Mouse primary spleen cells | PBMC | 1300 -1450 | NA |
Mouse bone marrow monocyte | PBMC | 1300 -1450 | NA |
Human Cultured T cells | Cell Line | 480-540 | 1150-1300 |
Human Cultured NK cells | Cell Line | 480-540 | 1200-1300 |
Human iPSC | Cell Line | 500-540 | 1200-1300 |
Human MM1.S | Cell Line | 520-560 | 1250-1350 |
Human adipose MSC | Cell Line | 500-540 | 1200-1300 |
Human bone marrow MSC | Cell Line | 500-540 | 1200-1300 |
Human cord blood MSC | Cell Line | 500-540 | 1200-1300 |
Human ESC derived neural stem cell | Cell Line | 520-560 | 1250-1350 |
Human primary hepatocyte | Cell Line | 480-520 | 1150-1250 |
Human primary Dendritic cells | Cell Line | 480-520 | 1150-1200 |
Human neural progenitor ReNCell VM | Cell Line | 520-560 | 1250-1350 |
Mouse iPSC | Cell Line | 480-520 | 1150-1250 |
Mouse ESC | Cell Line | 480-520 | 1150-1250 |
Mouse lymph node Cultured T cell | Cell Line | 500-540 | 1200-1300 |
Mouse peritoneal Macrophage | Cell Line | 540-580 | 1350-1400 |
Primary mouse cortical neurons | Cell Line | 520-560 | 1350-1400 |
Rat primary cortex neural progenitor cell | Cell Line | 520-560 | 1250-1350 |
293 | Cell Line | 440-480 | 1050-1150 |
293T | Cell Line | 460-500 | 1050-1150 |
A20 | Cell Line | 540-560 | 1250-1350 |
A549 | Cell Line | 540-560 | 1250-1350 |
ADSCs | Cell Line | 500-520 | 1200-1300 |
ARPE-19 | Cell Line | 520-540 | 1250-1350 |
BaF3 | Cell Line | 520-540 | 1250-1350 |
BJ | Cell Line | 500-520 | 1250-1400 |
CAL51 | Cell Line | 500-520 | 1250-1350 |
CHO-K1 | Cell Line | 560-600 | 1250-1400 |
CHO-S | Cell Line | 520-580 | 1250-1400 |
DEL | Cell Line | 500-540 | 1200-1300 |
DLD1 | Cell Line | 500-520 | 1200-1300 |
EMT6 | Cell Line | 500-520 | 1200-1300 |
HCT116 | Cell Line | 540-560 | 1200-1300 |
Hela | Cell Line | 480-520 | 1250-1350 |
Hepa1-6 | Cell Line | 500-520 | 1200-1300 |
Hep3b | Cell Line | 500-540 | 1100-1200 |
HSF | Cell Line | 520-540 | 1250-1350 |
HUC-Maaa13-P3 | Cell Line | 520-540 | 1250-1350 |
HUVEC | Cell Line | 520-560 | 1250-1350 |
H1975 | Cell Line | 520-560 | 1250-1350 |
H22 | Cell Line | 500-520 | 1200-1300 |
H226 | Cell Line | 520-540 | 1250-1350 |
H9C2 | Cell Line | 500-540 | 1350-1400 |
HL-60 * | Cell Line | 520-560 | 1250-1350 |
iMC | Cell Line | 500-540 | 1200-1300 |
Jurkat | Cell Line | 500-540 | 1200-1350 |
Karpas299 | Cell Line | 520-560 | 1250-1350 |
K562 | Cell Line | 480-520 | 1200-1300 |
MC38 | Cell Line | 500-520 | 1200-1300 |
MCF-7 | Cell Line | 500-520 | 1200-1300 |
MDA-MB-231 | Cell Line | 500-520 | 1220-1300 |
Molm-13 * | Cell Line | 500-540 | 1250-1350 |
Nalm-6 | Cell Line | 520-540 | 1250-1350 |
NK-92 | Cell Line | 440-460 | 1150-1200 |
Raji | Cell Line | 520-540 | 1250-1350 |
RAW264.7 | Cell Line | 520-540 | 1250-1350 |
SF9 insect cell | Cell Line | 480-520 | 1200-1300 |
SK-HEP-1 | Cell Line | 480-520 | 1220-1300 |
SP2/0 | Cell Line | 480-500 | 1150-1200 |
SUP-M2 * | Cell Line | 520-560 | 1250-1350 |
Swiss-3T3 | Cell Line | 520-540 | 1250-1350 |
p3X63Ag8.653 | Cell Line | 500-540 | 1220-1300 |
THP-1 * | Cell Line | 500-540 | 1250-1300 |
U251 | Cell Line | 480-520 | 1250-1400 |
U937 * | Cell Line | 500-540 | 1250-1300 |
*Many monocyte or macrophage related cell lines such as THP-1, U937, HL-60 and Molm13 will die out within two to three days after plasmid electroporation. These cells have no problem with mRNA, siRNA and Cas9 RNP electroporation. The efficiency would be nearly 100% and cell survival is not affected. Celetrix has developed the THP-1 Helper Kit for plasmid electroporation of monocytic cell lines. A significant portion of THP-1 and U937 cells can survive and grow with the helper kit, enabling easy generation of stable cell lines. Our THP-1 Helper Kit does not help cell lines such as Molm13 and SUP-M2 with plasmid electroporation and we are actively researching the solutions.
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