Christopher Willig

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Questions related from Christopher Willig

How to extract RNA from powdered plant tissue stored in RNAlater?

My research group has samples of plant tissue that were pulverized in ln2 using a mortar and pestle before being weighed into tubes and stored in RNAlater at -20 degrees for a few months before...

29 May 2024 7,307 0 View

Where can I get "Silwet-70" to use for plant transformation?

Hi, I want to buy some of the surfactant Silwet-70 to use for improving transformation frequency in cereal crops as described in publications at the following links: Developing a flexible,...

05 September 2020 820 4 View

What are some examples of plants with low transformation frequency (about 1-3% or lower) that have been edited with CRISPR?

Are there any literature references on gene editing or regular production of transgenic plants in species or genotypes that are very recalcitrant to transformation?

09 April 2020 6,909 0 View

Is it feasible to use CRISPR to induce bi-allelic long deletions (~100 kb) in plants?

Can this be accomplished using Cas9 and two sgRNAs targeting genomic sites over 100 kb away from each other? I want to do this in a plant that is transformable, but for which there are no reports...

05 March 2020 7,924 5 View

Can anyone recommend an E. coli strain other technique for cloning a repetitive DNA sequence?

I am trying to clone a promoter sequence using a pUC57 plasmid backbone in DH5alpha cells. The finished sequence will contain 6 direct repeats of a ~50 bp sequence I started from complimentary...

15 November 2018 431 6 View

What publicly available dataset(s) can I use as a proxy for estimating what proportion of a given genome is functionally characterized?

I want to be able, in some quantitative way, to compare several plant species with respect to how much is known about the functions of all known loci in their genomes. It seems like looking at...

17 October 2018 7,277 4 View

How can I perform a blunt-end ligation with a large vector and large insert?

I am trying to perform a rather difficult blunt-end ligation that requires several enzymatic steps and would appreciate any tips from those who have experience with blunt-end cloning. My...

07 June 2018 4,005 10 View

Should I digest with BsaI and gel purify my destination vector prior to setting up a Golden Gate reaction for cloning gRNA spacer sequences?

I'm planning to use Golden Gate cloning in order to insert guide RNA spacer sequences into a cloning vector between two head-to-head BsaI restriction sites. Prior to setting up the reaction with...

15 March 2018 4,853 1 View