Here's the latest from the Cloning & Expression Channel at Bitesize Bio.

Articles

10 ways to improve blunt-end ligations By Alex Reis Blunt-end cloning involves the ligation of DNA fragments – usually between a plasmid vector and an insert – whose terminal ends are not “sticky”. Performing these ligations is notoriously difficult, particularly with large DNA fragments. But it is possible. And in this article I’ll give you some tips that I hope will increase your chances of success ...[read more] Choosing the right plasmid vector: A Guide for beginners By Alex Reis f you’re picking up a list of available plasmid vectors for the first time, it can be mind-blowing to decide on the best one for your experiment. What cloning sites do you need? What restriction enzyme sites? What insert size is possible? Let me help you with some pointers about what factors to look out for.... [read more]

From The Archive

Using Enzymes at the Bench — Keep it in the cooler? On ice? or at RT? By Ellen Moran Have you ever needed an enzyme but not had an ice bucket? Have you been tempted to just grab the enzyme out of the freezer, take what you need and quickly return the tube to the freezer? Do enzymes really need to be kept on ice all of the time? I would say, Yes! ...[read more] Five Things You Might Not Know about LB By Ellen Moran LB medium is a staple in virtually every lab. It’s commonly used to propagate E. coli, and as such will be used frequently by any lab that does cloning. Chances are, LB broth or plates were one of the first things you learned to make as a newbie in the lab. Here are a few interesting facts you might not know about LB ...[read more] Use Less Vector, Killer Cut for Success in Plasmid Cloning By Nick Oswald Here’s an all-too-often repeated scene in the lab: First thing in the morning, you approach the 37°C incubator with trepidation, open it and through one half-open eye you take a look at the LB plate that you spread your ligation-reaction-transformed E.coli aliquot onto ...[read more] PCR Rescue: Making One Band From Many By John Mackay It’s the molecular biologist’s version of ‘I have good news. . and bad news’. The good news is that I amplified the DNA band of interest. The bad news is that I amplified these other bands as well! Oh, and this smear. What to do? ...[read more] DIY Electrocompetent E. coli By Nick Oswald If you buy competent E.coli regularly, you’ll know that they are pretty expensive. So the cost of screwing up a cloning or transformation experiment is pretty high in terms of money, as well as your time and sanity! ...[read more]