...: RNAi animation Whitehead siRNA Selection Program: http://sirna.wi.mit.edu/ Back to Top I. Appendix I...Below are suggestions for target selection. Use an siRNA selection tool to determine a set of top-scoring...Whitehead Institute for Biomedical Research hosts an siRNA Selection Program that can be accessed after a free... free registration. Guidelines for designing siRNAs with effective gene silencing: Starting at 25nt downstream.... References H.1. Published Articles Functional siRNAs and miRNAs exhibit strand bias . Khvorova A et....

...continue running the gel until you have obtained the desired separation. Gently open the gel with the spatula...Device. Close the lid of the device. Select the desired method and make sure the parameters are correct... incubation Dilute the primary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...peroxidase-conjugated secondary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...

...antibody and an isotype control antibody to the desired concentrations in the antibody dilution buffer....HRP-conjugated isotype-specific secondary antibody to the desired concentrations in the antibody dilution buffer....Check the plate periodically to determine when the desired color change has occurred. The color will change... to optimize experimental conditions. When the desired color change has occurred, gently remove the plate...

... per well. Allow the HeLa cells to grow to the desired density before labeling. Section 2: Fixing and ... container. Dilute the primary antibody to the desired concentration in antibody dilution buffer. Pro-...fluorescently-labeled secondary antibody to the desired concentration in antibody dilution buffer. Pro-...

..., but do not cut within your insert Are in the desired location in your recipient plasmid (usually in ... fear. You have other options, such as: Adding desired restriction sites to flank your insert : You can...restriction enzymes that cut within your insert. Adding desired restriction sites to your recipient plasmid : You...

...Spectrophotometer to see if the sample has reached the desired concentration. If the sample concentration is still...volume of the sample is reduced enough to reach the desired concentration. Pro-Tip Periodically check the concentration...Spectrophotometer to see if the sample has reached the desired concentration. Gently transfer the sample from ...

...Set up your water bath so that it will reach the desired temperature by the time you need it for your experiment...water bath to prevent evaporation and maintain the desired temperature. This also helps the water bath heat...

...-agar 1 L Sterile H 2 O Sterile plates of your desired size - we usually use 60 mm x 15 mm plates which...next to the flame and begin pouring. Measure your desired amount of agar with a pipete for the first plate...

...plasmid DNA, the plasmid will already be in your desired bacterial strain and you will not need to obtain...

...and with a clean, sterile razor blade, slice the desired DNA fragment from the gel. Notes: To protect the...

...you can design a set of oligos containing your desired restriction sites and add them to your existing...

...recheck the pH to prevent over or undershooting the desired pH. Allow the solution to mix for 10 min and then...

... likely to ligate to itself rather than to the desired insert. If you are in this situation, it is important...

...and yield. If there are significant amounts of undesired product, gel-purify DNA segments. Otherwise, PCR...

...recheck the pH to prevent over or undershooting the desired pH. Allow the solution to mix for 10 min and then...

...maximum possible setting on the pipette, set the desired volume of the liquid on the pipette. Depending ...

...dilutions and pick the population that has the most desirable level of expression. Over time, transgene expression...

...that: Do not cut within your insert. Are in the desired location in your recipient plasmid (usually in ...

...recheck the pH to prevent over or undershooting the desired pH. Allow the solution to mix for 10 min and then...