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Questions related from Md. Amzad Hossain
I have prepared competent cells of L. lactis NZ9000. But transformation not working. No colony at all in the plate. Parameters are as following: plasmid concentration: 1 uL (300ng) Electroporator-...
03 April 2024 1,148 3 View
I want to digest pET28a plasmid (5369 bp) with 2 restriction enzymes (RE)- EcoRI and XhoI for cloning. I checked the REs have single cutting site of each in the plasmid. However when I digest it...
11 June 2023 5,649 4 View
For cloning purposes, I have to double digest PCR products and plasmids 20 ug of each. Standard 50 uL restriction digestion reactions can accommodate up to 1 ug of DNA. According to some experts,...
23 May 2023 770 7 View
Hello Everyone I am trying to isolate pET28a plasmid (5.37 kbp) from E.coli strain. The electrophoresis result shows only one band that is >20 kbp. This is triplicated and same result comes all...
27 February 2023 2,330 4 View
I am planning to do Gibson assembly for the very first time. As per the recommendation of the assembly kit, 25-40 bp overlap is required between the plasmid and insert. I have designed a primer...
10 January 2023 8,323 3 View
Cloning a 6.5 kbp DNA in an expression vector failed using the heat shock method. Are there any troubleshooting points in this case?
15 November 2022 7,320 1 View
I am working with a gram positive bacteria. Please suggest a protocol for good yield.
27 October 2022 4,424 3 View
I need to transform a plasmid of 10 kbp in Gram positive competent cell (L. cremoris NZ9000). Right now, I do not have electroporation facility. Is there any successful record of heat shock...
01 January 1970 3,268 0 View
