| Autor: |
Zeinab Sharafi, Sina Adrangi |
| Jazyk: |
angličtina |
| Rok vydání: |
2019 |
| Předmět: |
|
| Zdroj: |
Trends in Peptide and Protein Sciences, Vol 3, Pp 1-8 (e1) (2019) |
| Druh dokumentu: |
article |
| ISSN: |
2538-2446 |
| DOI: |
10.22037/tpps.v3i0.19997 |
| Popis: |
Ligation-independent cloning is a simple method that provides several advantages over conventional cloning. However, the efficiency of ligation-independent cloning is considerably lower than that of conventional methods. Several studies have shown that competent cells used for ligation-independent cloning should preferably have a transformation efficiency of 106-107 cfu/μg DNA. Although such levels can be easily achieved using standard protocols with most Escherichia coli strains, some strains attain mush lower values. When such strains have to be used for ligation-independent cloning, certain measures need to be taken to avoid any situation that may further decrease the efficiency of the process. These measures, however, are usually time-consuming. This problem is exacerbated by the fact that some strains such as BL21 (DE3) appear to be intrinsically unsuited for ligation-independent cloning. Here we suggest that by avoiding DNA damage during purification simply by replacing UV transilluminators with blue light systems BL21 (DE3) cells with a transformation efficiency of 105 cfu/μg DNA can satisfactorily be used for ligation-independent cloning without any additional steps. HIGHLIGHTS •Using blue light instead of UV light increases the efficiency of LIC. •A simple LED blue light projector combined with a filter can be used for this purpose. •This approach is not recommended for applications that require higher sensitivity. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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