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... that increase the transformation efficiency and make bacteria more susceptible to either chemical or ... plasmids at once. To save money, many labs also make their own competent cells. This is a relatively ...

...endonuclease negative E. coli strain such as NEB stable. Make a mixture of a total of 500 μL PEI-OptiPro SFM with...-Tip There can be batch to batch variation when making the PEI working stock, therefore the ratio of μg...

...plunger. Remove the entire pipette from the container making sure not to touch the sides of the container with...important for scientists to calibrate their pipettes to make sure that they are properly tuned to dispense the...

...solutions at various dilution. Make sure to use the polybrene-containing media to make the cell solution in this...

...In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding...restriction site (GAATTC) to the 5’ end of this primer, making our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC...

...buffer. Fill the chamber with 1X MOPS running buffer. Make sure that the chamber is sealed. Load 5 µL of the...Pro-Tip Leaving clear lanes between samples will make quantifying the gel easier. Cover the electrophoresis...

...bottle up/down vigorously for 30 sec (it’s okay to make bubbles). Incubate at RT for 15 min. Note that longer...add the OptiMEM + DNA + PEI solution to the CS5. Make sure that all five layers are covered with media...

.... See below for options if the recombinant band makes up a significant proportion of the DNA pool. Last... weight of each pellet should be ~1-2 g. Pro-Tip Make sure to weigh the empty tube beforehand! If you’...

... and 3' - G and CAGCT - 5' to the bottom oligo, making our final oligos 34 bp each: Top oligo: 5' - AATTC...

...appropriate water level in the water bath so before use, make sure that there is enough water inside....

...sites, will result in similar sized bands, thus making this simple digest less informative. This is particularly...

...new window) Video Link Description Related Page Making LB Agar Plates Create plates to culture bacteria...

...Description (Link opens in a new window) Link to Video Making LB Agar Plates Create plates to culture bacteria...

...instructions. If your enzyme did not cut, check to make sure that it isn't methylation sensitive . ...

...lid of the device. Select the desired method and make sure the parameters are correct. Transfer for 5–...