Essay on problems occuring during ligation

Arnold

New member
Nov 16, 2015
1
0
0
2
#1
Hello All,



I'm a Bachelor student Biomedical Sciences and working on an essay on problems wich occurs during cloning in a vector. I'm looking for some reviews on problems wich occurs during cloning in a plasmid/vector? Do you know some usefull articles and reviews? At the moment I found the most questions on ligation problems on fora. Some causes/subjects I summed up below. Anyone now more common mistakes and problems during digestion of an insert or some articles/reviews with solutions on these mistakes/problems?

[list type=decimal]
[*]There is a second same restriction site in the vector.
[*]Sticky ends not complementary
[*]Low digestion rate because of blind ends restriction enzymes
[*]Enzymes are not working or wrong temperature.
[*]Vector-vector ligation (solution is to dephosphorylating the ends using SAP)
[*]Ratio DNA:plasmide
[/list type=decimal]
Quantity of DNA is not of interest (length), but the numbers of inserts are important.

Using two differenct restriction sites is important to guarantees the orientation of the insert

3:1 insert to vector?



Thanks a lot,



Arnold



 
1

1

Guest
#2

With such website you may be able to get the cheat pokemon go online and there can be instant free pokecoins in your account.