Md. Amzad Hossain

6 Questions 5 Answers 0 Followers

Questions related from Md. Amzad Hossain

What are the reasons of unusual digestion pattern of EcoRI?

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 5,646 4 View

Can anyone suggest a 10 ug restriction digestion reaction recipe?

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 734 7 View

Is it possible to get only multiple plasmid band in gel?

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 2,324 4 View

How to design primers for Gibson assembly not forming secondary structure?

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 8,319 3 View

Does heat shock method of transformation work to clone 6.5 kbp DNA?

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 7,314 1 View

Can anybody suggest a successful protocol of bacterial genomic DNA isolation?

I am working with a gram positive bacteria. Please suggest a protocol for good yield.

27 October 2022 4,420 3 View