...first gel lane (as in Figure 2). Select Analyze . Select Gels . Select Select First Lane . Drag the box to...to the next lane. Select Analyze . Select Gels . Select Select Next Lane . Repeat for all lanes. Plot ...protein bands as follows: Figure 3 Select Analyze . Select Gels . Select Plot Lanes . One graph per lane ...the Chart Editor , select the Customize tab. Select the Series drop down menu. Select the Trendline checkbox... lane. Import the gel image into ImageJ. Select File . Select Open . Choose the location of the file to... open. Change the image type to 8-bit . Select Image . Select Type . Choose 8-bit . Determine each lane...the bottom of each peak. Select the wand tool. Fill in each peak by selecting the center of the peak. ...

...for effective over-agar selection has been empirically determined. See selection curve below. Carbenicillin..., plated over-agar for selection of transformed E. coli . Sample Data Selection of E.coli on LB-agar using...1 mg/mL and 2 mg/mL plates and effective selection. Selection Curve of Transformed E. coli after Over-...methodology for plating antibiotic over-agar for the selection of transformed E. coli . Over-agar spreading of...easy for an investigator to conveniently plate and select transformed cells containing plasmids differing...agar. This protocol will focus specifically on selection with carbenicillin; however it can be adapted ...adapted for any antibiotic through the use of a selection curve to determine optimal antibiotic concentration...

... target selection are continuously being improved. Below are suggestions for target selection. Use an ...concentration F.3 Protocol for lentiviral infection and selection G. Safety H. References H.1 Published articles...encoded in pLKO.1 allows for convenient stable selection. Figure 1 : Map of pLKO.1 containing an shRNA ...puromycin. Puro R Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian cells. sin 3’LTR...replication. Amp R Ampicillin resistance gene for selection of pLKO.1 plasmid in bacterial cells pUC ori pUC...Determining the Optimal 21-mer Targets in your Gene Selection of suitable 21-mer targets in your gene is the...an siRNA selection tool to determine a set of top-scoring targets for your gene. For example, the Whitehead...

...enables selection of a stable cell culture after transduction. Performing antibiotic selection on transduced...This is the beginning of the selection process, which will begin the selection of a stable cell pool. Observe...transducibility of the cell line used, this antibiotic selection may be a vital step for obtaining a population...Remove media, replace with fresh media containing selection reagent Day 3–14: Change media as needed Day 14...Lentivirus preparation Appropriate antibiotic for selection (e.g. puromycin, blasticidin) Reagent Preparation...Before beginning, determine the optimal dose of selective reagent for your target cell line. To do this,...resulting in massive cell death upon antibiotic selection. The number of cells transduced should be enough...

... thought on using selective medium throughout subsequent experiments. When selective medium is used, it...was used for the initial selection). Addgene recommends that the use of selective medium throughout experiments...site(s) of proviral integration across cells. Selective pressure on this heterogeneous cell pool could...the polyclonal cell pool is always undergoing selection, it’s best to generate monoclonal cell lines from... sure that the cell pool has been sufficiently selected with the appropriate antibiotic so that every ...biological effects on the cell line. In the absence of selective medium, there is a chance that the integrated ... Some researchers choose to maintain cells in selective medium at a reduced antibiotic concentration (...

... of selective reagent for 12 days with media exchanges every 3–4 days. After 12 days of selection, no ...adapted to a variety of target cell lines and selection markers. Note this assay requires staining resistant... 2: Replace media with fresh media containing selection reagent Days 3–14: Change media as needed Days...Colonies were allowed to grow in the presence of selective reagent for 12 days with media exchanges every...

...Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and how to handle...experimental results. This protocol will guide you in selecting the correct pipette and pipette tip and using ...know the different parts of the pipette, how to select tips, and how to read a pipette, you’re ready to...display or the top of the plunger of the pipette. Select the pipette tip that is designed to fit your pipette...

...bacteria that do not contain it by artificial selection (i.e. growing the bacteria in the presence of ...addition of an antibiotic to this gel allows for the selection of only those bacteria with resistance to that...stable, so it is potentially more effective at selecting only bacteria containing the plasmids of interest...the antibiotic at too low a concentration for selection. Negative Result 2: Neither Strains Grows If neither...

...most widely used method scientists employ to selectively move a specific piece of DNA from one plasmid...plasmid contains the expected size insert. By selecting the appropriate enzyme(s), one can either linearize...beginning your diagnostic digest, you will need to select a restriction enzyme or enzymes that cut your plasmid...

...for a wide range of applications, with videos for select protocols in the right-hand column. You can find...and use Watch the Video! Pipetting Learn about selecting the correct pipettor and pipette tip, how to dispense...pipette Watch the Video! Centrifugation Learn about selecting and using a centrifuge to separate different components...

...same technique can be used to move promoters, selectable markers, or any other DNA element between plasmids...restriction sites are present in a given sequence. When selecting restriction enzymes, you want to choose enzymes...information about restriction enzyme buffers). If you select enzymes that can function in the same buffer, it...

...min for proteins >150 kDa. Select Start Run. When the run is complete, select Done. Section 5: Blocking...Gel Transfer Device. Close the lid of the device. Select the desired method and make sure the parameters...

...high-throughput next generation sequencing (NGS). Select a restriction enzyme for digest that will cut the...propagation (origin of replication and antibiotic selection). This recombination, at a low rate, is not typically...Recombination is thought to be in part related to selective pressure. These recombined plasmids are much smaller...

...proteins, point mutations, deletions, etc.) involves selecting one or more unique oligonucleotide primers that...There are a number of factors to consider when selecting sequencing primers, including: Is the primer unique...

...bacteria that do not contain it by artificial selection (i.e. growing the bacteria in the presence of ...flask. Using a sterile pipette tip or toothpick, select a single colony from your LB agar plate . Drop ...

...the reaction mixture, and may provide counter-selection for negative clones as in the example shown here...removing the stuffer sequence and sacB counter-selection marker (see our restriction digest protocol )....

...sequence to be amplified (usually 18-21bp). When selecting restriction sites, you should use a DNA analysis... longer the expected PCR product is. You should select an annealing temperature based on the melting temperature...

...for 1 hour. Lithium Chloride (Option B): LiCl selectively precipitates RNA versus DNA or proteins. Add ...

...chemically by joining nucleotides together. One must selectively block and unblock repeatedly the reactive groups...

...Learn about selecting and using a centrifuge to separate different components in a liquid sample....