...in size. The structure of the primer should be relatively simple and contain no internal secondary structure...

... your experiment. Water baths tend to heat up relatively slowly. Place the water bath cover on the top...

... Let's assume that your favorite vector has a relatively limited MCS (BamHI - EcoRI - SalI) and you want...

...protocol. Pro-Tip Spin speed is often given as Relative Centrifugal Force (RCF) or Revolutions Per Minute...

... across the dilutions. RPP30 copies should be relatively constant across samples. In the RRE example below...concentration of RPP30 positive droplets stays relatively even across samples (green). To increase the ...

...also make their own competent cells. This is a relatively simple procedure and is useful for performing...idea to use electro-competent cells. Instead of relying on the heat-shock to cause the cells to take up...

...be in the liquid. Pro-Tip If pipetting from a relatively large container holding a small volume of liquid...samples. Tips can come loose in a bag, or can come preloaded into tip boxes. In either case, once assembled.... Once the end of the tip is submerged, slowly release the plunger, allowing it to go back to its resting... the plunger all the way to the second stop to release the liquid. Pro-Tip Dispense until the first stop... expel any residual liquid in the tip. Before releasing the plunger, lift the pipette tip out of the liquid...Once you are clear of the liquid you can slowly release the plunger. Remove the entire pipette from the...

...be 10 µg/mL. Pro-Tip Transducing too many cells relative to the number of virus particles reduces the transduction...antibiotic in culture, or remove the antibiotic entirely. If the antibiotic is reduced or removed from ...

...) or a similar photo software to determine the relative intensity of the protein bands to the overall ...

...hemocytometer or other cell counter. Pro-Tip Prepare a relatively dilute cell solution (<10 6 cells/mL) before ...

... A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B...TRC cloning vector has a 1.9kb stuffer that is released by digestion with AgeI and EcoRI. shRNA oligos...terminal repeat. RRE Rev response element. A.3 Related Products The following plasmids available from ... have at least 3 nucleotide mismatches to all unrelated genes. TIP: Addgene recommends that you select...cloning vector contains a 1.9kb stuffer that is released upon digestion with EcoRI and AgeI. The oligos...

...reference (typically ROX dye), to normalize non-PCR–related fluorescence fluctuations and to minimize well-...tested. Plasmid #59462 is one plasmid that gave reliable and consistent results. Use the recommended plasmid...following: Standard curve: R 2 (coefficient of correlation) ~ 1.0, E (efficiency of PCR) ~100% (90%–110%...

...purification. Immediately purify them! Commercial Maxipreps rely on incremental, ordered cell lysis. E. coli cells...digest. Recombination is thought to be in part related to selective pressure. These recombined plasmids...can dramatically plateau and sometimes fall off entirely if the column or reagents are significantly overloaded...

...beginning or end of a sequencing reaction is often unreliable. Although your sequencing results may indicate... file, you will see that these base calls are unreliable. This is an example of a trace file from a portion...

... opens in a new window) Video Link Description Related Page Making LB Agar Plates Create plates to culture... opens in a new window) Video Link Description Related Page MTA Guide - Requests Your guide to the MTA...

...impenetrable by a needle, and must be able to close securely. Biohazard signs should be placed in all areas...

... Note: This step causes the bacteria to lyse, releasing their contents, including plasmid DNA, into solution...