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RNA Extraction Without A Kit

Posted by Leah Schwiesow on Mar 24, 2020 9:15:00 AM

As with DNA isolation, scientists commonly rely on RNA isolation kits to make their life easier. Recently, we published a blog on DNA purification without a kit that outlined several reasons why doing something without a kit has advantages: less plastic waste, less expense, and less of being left with a bunch of random solutions when all the spin columns run out. In this article, we cover the basics of isolating RNA without a kit.

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Topics: Molecular Biology Protocols and Tips, Plasmids

DNA Purification Without a Kit

Posted by Beth Kenkel on Feb 11, 2020 9:15:00 AM

Before you reach for that silica spin column, stop to consider some ways to purify DNA without a kit. DNA purification kits have advantages: they are convenient and provide uniform, consistent results. But they are also less accessible due to their expense and requirement for lab equipment. Plus they create plastic waste. Kits can also have the annoying tendency to runout right when you need them and to accumulate a bunch of unused buffers because you’ve run out of columns.

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Topics: Molecular Biology Protocols and Tips, Plasmids

Harnessing Bacterial Toxins for Allelic Exchange

Posted by Guest Blogger on Aug 15, 2019 8:30:02 AM

This post was contributed by Jacob Lazarus, a postdoctoral researcher at Harvard.

There’s an astounding number of ways to create chromosomal mutations in bacteria, so many that it may be difficult to decide which path to take. A quick and easy way to introduce a mutation in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes. However, there are ways to create scarless mutations, ones that don’t leave any undesired scars in the chromosome.

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Topics: Molecular Biology Protocols and Tips, Other Plasmid Tools, Plasmids

Save Time with Transient Plant Leaf Transformations

Posted by Guest Blogger on Jul 25, 2019 8:03:52 AM

This post was contributed by Samuel Mortensen, a PhD candidate at Northeastern University.

Working with plants doesn’t always have to be a time-consuming process. While developing transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant can take even longer, a transient plant leaf transformation process could save the plant biologist some time… months, in fact.

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Topics: Plant Biology, Molecular Biology Protocols and Tips, Other

Tips for arabidopsis transformation

Posted by Guest Blogger on Oct 25, 2018 9:23:48 AM

This post was contributed by Laura Lee, a graduate student at Stanford University.

Arabidopsis is a fantastic model organism for many reasons, not the least of which is ease of transformation. There are many motivations to generate transgenic Arabidopsis, from studying transcriptional and translational dynamics of genes and proteins in living plants, to complementing mutant phenotypes. Arabidopsis is amenable to the floral drip or dip transformation method. The general steps for this method include:

  • Cloning and transforming a plasmid into the bacterium Agrobacterium tumeficans - a plant pathogenic species that stably integrates transfer DNA (tDNA) into the genomes of the plants it attacks
  • Growing the transformed agrobacterium culture
  • Dipping your plant’s flowers in the agrobacterium culture to allow for tDNA insertions into the plant’s germline
  • Selecting for seeds that have the tDNA insertions (usually via seed growth on antibiotic-containing media)
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Topics: Plant Biology, Molecular Biology Protocols and Tips, Plasmids

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