반응 #305473

반응 방정식

G418

neutral red

crystal violet

→

Neutral Red Crystal Violet

반응 조건

후처리

1. 1
   기타The medium was removed from the cells
2. 2
   workup.ADDITIONcontaining 0.0025% neutral red (Sigma N2889)
3. 3
   세척Cells were washed twice with PBS
4. 4
   workup.WAITfixed in 3.5% formaldehyde for 15 minutes
5. 5
   세척washed twice again in PBS
6. 6
   workup.WAITby incubating with an 0.1% crystal violet (Sigma C0775) solution in 20% methanol for 20 minutes at room temperature
7. 7
   세척in 20% methanol and a wash with distilled water

실험 절차

The survival of transfected cells in the absence or presence of G418 was monitored by staining of foci/clones with neutral red in vivo with subsequent crystal violet staining. The medium was removed from the cells and replaced with fresh medium containing 0.0025% neutral red (Sigma N2889) and the cells incubated for 3 hours at 37° C. Cells were washed twice with PBS, fixed in 3.5% formaldehyde for 15 minutes, washed twice again in PBS and then with distilled water and the number of (live) foci counted. The cells could then be re-stained with crystal violet by incubating with an 0.1% crystal violet (Sigma C0775) solution in 20% methanol for 20 minutes at room temperature, followed by 3 washes in 20% methanol and a wash with distilled water.

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