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2

Addition to what Chris already said: Papain can be used for cells sensitive to trypsin (neurons etc) Collagenase can be used for certain cells where trypsin is ineffective (Accutase is a commercially available enzyme mix(?) which has collagenase activity) Hyaluronidase - I don't know where it is specifically preferred but it is used. Pronase and ...


4

Lets make this a proper answer: There are a few possibilities to detach adherent cells without Trypsin. PBS/EDTA: Integrins and Cadherins play an important role in the adhesion and also in maintaining cell-cell contacts. These function of these proteins depends on Calcium2+ ions, so EDTA will chelate them and make them unavailable. First remove the culture ...


1

That is not true. If you had forgotten to add the antibiotic before inoculation then you can add it before the bacteria starts growing. Make sure you add it when the bacteria is still in lag phase. If you add it later then it won't be effective as some non-resistant (non-transformed) bacteria would have already expanded their population. Ampicillin and ...


2

I hate this problem but I hate even more when I pull up a cryo vial that some one wrote on with a dry erase marker. Get printable tough tags. Two options after that, print on them with your laser printer. Can do many at a time. You can take a step further and overlay the tag with some 3m single sided tape (scotch tape is gonna brittle up from the N2.) ...


5

This is indeed a problematic point. I habe been using machine printed labels for a while now (sometimes with some extra scotch tape around it to prevent it from falling) off, which worked pretty nicely. However, there is a printer available on the market (though I haven't tested it yet) which claims that they can print on any tube. This printer is called ...


3

What I find works is to scotch tape the labels to the tubes, they hold up fine in our -70C freezer. What I did was go into excel 2010, go to View tab, click on Page Layout on the left side, not normal view! Then, go to home tab, select all the cells on the page. Go to the right side of the home tab, hit Format, then Row Height. This lets you set height in ...


6

You have two possibilities: When you only need a quick check if your RNA is ok and you indeed only get one band, you can try a "quick and dirty" method. Heat the sample for 5 minutes at 65°C and then immediately cool it down in an icebath and keep it there until loading. By doing so, you melt up the secondary structure of the RNA and keep it in this state. ...


2

RNA is amenable to folding, which produces secondary structure and alters its mobility (compare to supercoiled DNA). Therefore, if you don't denature your RNA appropriately, even a single species will produce a smear. A common strategy is to incorporate denaturing chemicals such as formaldehyde into the procedure. See this Life Technologies protocol. ...


7

Wear two sets of gloves. First, use nitrile gloves, not latex for this. 4mm or thicker is a specification I would recommend to avoid tearing. Latex will bind too well to itself and tear easily with increased friction etc (same logic behind not using two condoms). Also, use a coat with an elastic cuff on the sleeve. So here's the breakdown Put on first ...


2

Along with @shigeta's answer, I'd highly recommend Springer/Humana Press's series Methods in Molecular Biology, which started out as just a single collection of protocols (as I recall...) and is now a series of 1208 books, published between 1984 and 2015. There are a ton of protocols available online at places like Springer Protocols and Cold Spring Harbor ...


3

Molecular Cloning is updated and in its 4th edition. Every lab used to have it. Its comprehensive, but really no book could be complete.



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