...purification kit, such as the (Link opens in a new window) QIAquick Gel Extraction Kit . Always follow the...next intended step with the now gel purified DNA. Find more information about DNA quantification here ....in 10μL, make 1.1μg in 11μL. Reference Page Top Index...

...Find step-by-step instructions on how to use a water bath in the lab....different sizes in the lab. Some water baths contain individual slots for heating up tubes while other water ...manual. If you’re filling up a water bath with individual slots for tubes, remember that when you add the...

...LB agar plates are frequently used to isolate individual (clonal) colonies of bacteria carrying a specific...copy number refers to the number of copies of an individual plasmid within a single bacterial cell. Large...obtain a higher cell density. Reference Page Top Index...

...cover the general procedure and features to keep in mind when using any centrifuge. However, each centrifuge...like the G Force calculator (Link opens in a new window) . The centrifuge should come up to speed with ...centrifuge begins to wobble or shake, this could indicate an unbalanced or improperly loaded centrifuge....

...post transfection in individual harvests or a combined harvest where all the individual harvests are pooled...transfection. Test a variety of brands and lots of FBS to find one suitable with your protocols. FBS can be purchased...

...dispensed. The boxes that the tips come in often indicate a volume range that the tip can hold. This should...pipette you’re using, you will be rotating dials behind the numerical display or the top of the plunger...no liquid should drip from the tip. This could indicate that the tip is not on the pipette properly. Place...

...recommended plasmid, or test multiple plasmids to find a suitable one. Some labs have reported better results...second peak at a temperature of ~70–75 °C usually indicates the presence of primer dimers which can increase...Methods. 2012 Feb;23(1):18-28. (Link opens in a new window) PMID: 22428977...

... Chambers Cell-Stack (CS5) (Link opens in a new window) (3,180 cm 2 - the same surface area as 21 x T-...transfection. Test a variety of brands and lots of FBS to find one suitable for your protocols. FBS can be purchased...1.8-fold ( Rego et al., 2018 (Link opens in a new window) ). To prepare D1 + 0.1 M sorbitol, add 57 mL of...

...plasmid DNA from it, you will want to isolate an individual clonal population (single colony) of bacteria... proceed to Recovering Plasmid DNA or use the individual colonies for other experiments....

...NEB’s online tool NEBcloner (Link opens in a new window) , to help guide your reaction buffer selection...errors may occur. Addgene makes no warranty of any kind regarding the contents of any literature. Literature...IS” “AS AVAILABLE” basis without warranty of any kind either expressed or implied, including but not limited...

...anneal to a specific region of the insert. You can find Addgene's sequencing results by clicking on the ...unreliable. Although your sequencing results may indicate bases at specific locations, by looking at the...

... . The pattern of the fragments on the gel can indicate if the plasmid contains the expected size insert... or restriction enzyme -based cloning to test individual clones before use of more expensive forms of ...

...transfection. Test a variety of brands and lots of FBS to find one suitable with your protocols. FBS can be purchased...Check the cells for mycoplasma (Link opens in a new window) regularly Do not over- or under-grow your cells...

...4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI) Bovine serum albumin (BSA) Triton X-100...may need to try a variety of fixation methods to find the best conditions for your target. The optimal...

... μl of lab stock solution per 100 mL gel). EtBr binds to the DNA and allows you to visualize the DNA under...10 μL, make 1.1 μg in 11 μL. Reference Page Top Index...

...mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been processed correctly... Image Analysis Use ImageJ (Link opens in a new window) or a similar photo software to determine the relative...

... 2 h at RT but may result in more non-specific binding. Wash the membrane 3x for 5 min in 1X TBST at RT...You may need to try a variety of lysis buffers to find the best for your target. The optimal concentration...

...opens in a new window) Chomczynski P and Sacchi N, 2006 and (Link opens in a new window) TRIzol® User ...

...the next dilution. Repeat for all dilutions as indicated in the chart below. Dilution Factor Virus Volume...hgtb.2017.198. PMID: 29378428 (Link opens in a new window) ....

...how to design primers for PCR. Reference Page Top Index...