No discussions found
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
-After cell harvesting, chromatin must be sheared to 200 to 600 bp before ChIP.
-Optimize shearing conditions for your specific cell type and fixation protocol before starting a ChIP. Therefore, start with a small sample (1x 10e5 to 1x 10e6 cells) and check the shearing efficiency. |
-Keep the beads homogenously in suspension at all times when pipetting. Variation in the amount of beads will lead to lower reproducibility.
-Make sure that there are no crystals in the Buffer B. Gently heat and mix until crystals disappear.
- Chromatin from 1,000 cells is sufficient for one ChIP assay. |
-For longer term storage, aliquot 130 µl of sheared chromatin into cryotubes, snap-freeze in liquid nitrogen, and store at -80°C. |
| Upstream tips |
-After cell harvesting, chromatin must be sheared to 200 to 600 bp before ChIP.
-Optimize shearing conditions for your specific cell type and fixation protocol before starting a ChIP. Therefore, start with a small sample (1x 10e5 to 1x 10e6 cells) and check the shearing efficiency. |
| Protocol tips |
-Keep the beads homogenously in suspension at all times when pipetting. Variation in the amount of beads will lead to lower reproducibility.
-Make sure that there are no crystals in the Buffer B. Gently heat and mix until crystals disappear.
- Chromatin from 1,000 cells is sufficient for one ChIP assay. |
| Downstream tips |
| -For longer term storage, aliquot 130 µl of sheared chromatin into cryotubes, snap-freeze in liquid nitrogen, and store at -80°C. |
| Upstream tips |
Protocol tips |
Downstream tips |
-If any reagent has formed a precipitate, warm at 55 °C until dissolved. Allow to equilibrate to room temperature before use.
|
-Use 1 µg of antibody per well.
-The range of cells that can be used with this kit is 0.2–2.5 x 105/well/ChIP sample. Using more than 0.5 million cells per well will increase the non-specific binding and reduce the yield and specificity of the ChIP reaction. If higher yield of ChIP DNA is desired for downstream applications (e.g. ChIP-Seq., ChIP-chip): a) DNA could be pooled by eluting consecutively (with 50 µl of elution buffer) from multiple individual ChIP reactions. |
|
| Upstream tips |
-If any reagent has formed a precipitate, warm at 55 °C until dissolved. Allow to equilibrate to room temperature before use.
|
| Protocol tips |
-Use 1 µg of antibody per well.
-The range of cells that can be used with this kit is 0.2–2.5 x 105/well/ChIP sample. Using more than 0.5 million cells per well will increase the non-specific binding and reduce the yield and specificity of the ChIP reaction. If higher yield of ChIP DNA is desired for downstream applications (e.g. ChIP-Seq., ChIP-chip): a) DNA could be pooled by eluting consecutively (with 50 µl of elution buffer) from multiple individual ChIP reactions. |
| Upstream tips |
Protocol tips |
Downstream tips |
|
-Always cap spin columns before placing them in the microcentrifuge.
-Ensure that all buffers are in clear solution. Shake or vortex if these buffers precipitate.
-The conditions of cross-linked DNA shearing can be optimized based on cells and sonicator equipment.
If desired, remove 5 µl of sonicated cell lysate for agarose gel analysis. |
|
| Protocol tips |
-Always cap spin columns before placing them in the microcentrifuge.
-Ensure that all buffers are in clear solution. Shake or vortex if these buffers precipitate.
-The conditions of cross-linked DNA shearing can be optimized based on cells and sonicator equipment.
If desired, remove 5 µl of sonicated cell lysate for agarose gel analysis. |
Can't find the product you've used to perform this experiment? It would be great if you can help us by
Adding a product!