Tagged Questions
1
vote
0answers
24 views
Experience with Detox Cloning
I just saw this paper.
A Highly Efficient Molecular Cloning Platform that Utilises a Small Bacterial Toxin Gene
I was curious if anyone had experience with this cloning strategy using the IbsC ...
2
votes
3answers
82 views
Improving Gel Extraction yields
How can I improve my Gel Extraction yields. We use the standard protocol from Qiagen, gel extraction, dissolve in QG buffer at 42C and purify via anion exchange columns. However, with 500 ng we ...
2
votes
0answers
22 views
How does LCR compare to Assembly PCR
The question pretty much explains itself. How do the two methods compare? I've always used Assembly PCR but the method is prone to mistakes and I'm curious how it compares to Ligase Chain Reaction ...
6
votes
1answer
365 views
Does anyone have any TOPO directional cloning tips? [closed]
I'm just about to start working on a TOPO cloning after I couldn't get it to work with standard restriction/ligation. Does anyone have any tips for TOPO cloning?
5
votes
2answers
1k views
Primer design for introduction of restriction sites flanking a gene of interest
I am wondering what the correct method for primer design to introduce restriction sites. Specifically between two methods.
1) Primer first partially hybridises to the gene, has a mis-match where the ...
10
votes
2answers
187 views
What is the highest competency possible for E coli?
I am looking to find a highly competent E coli strain. I am making a library of a ~6.6kb plasmid and I am not getting high enough efficiency. Does anyone have a suggestion of a strain/protocol with ...
7
votes
2answers
515 views
How do I prepare and clone from E. coli DNA?
I'm looking for a protocol to get genomic DNA from an E. coli sample so that I can clone a small portion of it using PCR into a plasmid. (< 500 bp in this case).
It seems OWW (Open Wet Ware) ...
