SNAPGENE VIEWER GEL SERIAL
To plate these samples, we used the serial dilution technique.For each collection site, we labeled 3 nutrient agar plates as follows: “ direct count,” “ 1:10,” & “ 1:100“.Collection and Sampling of Environmental Samples at Environmental Education Center.Draft genome sequence of Janthinobacterium lividum strain MTR reveals its mechanism of capnophilic behavior. Valdes, N., Soto, P., Cottet, L., Alarcon, P., Gonzalez, A., Castillo, A., … Tello, M.
SNAPGENE VIEWER GEL SKIN
Janthinobacterium lividum colonizes the skin of some amphibians and confers protection against fungal pathogens. Janthinobacterium lividum is a rod-shaped, Gram-negative, motile, aerobic bacterium, commonly isolated from of soil and water of cold regions.
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Also, a free version of SnapGene, SnapGene Viewer, will read these files. The GCK files require Gene Construction Kit from Textco to view. Use a different pSIM plasmid, such as pSIM29, if you are targeting with an AmpR cassette.īelow are sequence files of the plasmids and other useful sequences in three formats, “pdf”, “text” or GCK. This can cause unexpected recombinants if you are targeting an amp-cassette into cells containing this plasmid. ** Be aware that these plasmids have homology to the AmpR cassette, sequence flanking the drug resistance gene but are not AmpR. Template for tetA, kan, amp, cat, and tetA-sacB cassettes, amplify with colony PCR. W3110 araD tetA-sacB-amp fliC cat argG:: Tn5 Oligo or dsDNA recombination with plasmids. High frequency oligo recombination when targeting plasmids. The beta gene is fused to the start codon of cIII.ĭefective for MMR thus gives high frequency oligo recombination. However, recombination with dsDNA is reduced ~10-fold. Gene and thus avoids toxicity associated with it. Defective for MMR (methyl-directed mismatch repair). Will not work for dsDNA recombination as it has no exo It is a useful control to verify that your oligo recombination is working. This strain contains the galK assay system for oligo recombination. Useful for moving prophage into other genetic backgrounds by P1 transduction using linked Tn10. The list of plasmids is below the strain list. Thanks!įor your convenience, the below strain list with complete genotypes can be downloaded as a Word file by clicking Recombineering Strains.docx. If you are a company and would like some of our strains, we must have you contact Rose Freel ( first at the NIH Office of Technology Transfer. With your request, please include your FEDEX number, your phone number and your address to expedite shipping. Strains and plasmids can be requested by sending us an email. If you want strains specifically for doing BAC work, see this site first. Maintaining our strain collection is time consuming and expensive for us. To date, we have sent out more than 5000 orders for strains and plasmids! Ask for them by strain or plasmid number, but please ask only for what you really need. You can mini-prep plasmid DNA and then transform them into your bacterial strain of choice to enable recombineering. We are happy to send you strains to get recombineering working in your lab. We also are happy to send you plasmids as transformants. "Recombinase"-independent recombineering.FAQ 14: Why don’t I get the high frequencies that you report for ssDNA oligo recombineering?.FAQ 13: What do I do with the tube containing the “gel-like stuff” in it when you send me a strain or plasmid?.FAQ 12: How should I maintain the recombineering strains?.FAQ 11: Is the amount of NaCl in the LB medium important?.FAQ 10: Why do my electroporations “pop” (arc)?.FAQ 9: What conditions should I use for electroporation?.FAQ 8: I get tons of recombinants but when I analyze them, none are in the right place.FAQ 7: Why don’t I get any recombinants?.FAQ 6: What happens if I induce for longer or shorter times?.FAQ 5: Do the oligos I order need to be purified?.FAQ 4: Can’t I use an air shaker for the 42° induction?.FAQ 3: What happens if I grow the recombineering cells at temperatures higher than 32☌?.FAQ 2: What sequences do I use to make a drugR cassette for knock-outs?.FAQ 1: How much antibiotic should I use for chromosomal/BAC (single-copy) or plasmid (multi-copy) constructs?.“Recombinase”-independent recombineering.On the mechanism of ssDNA recombination.
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