What are the best conditions to improve transformation efficiency of XL1 Blue competent cells?
Please suggest any other conditions that you use to troubleshoot transformation efficiency in XL1 Blue competent cells.
Dear Vikas Ranvir
one of the Catalog attached: Number 200138 Product Name XL1-Blue MRF’ Kan Library Pack Competent Cells.
Competent cells are sensitive to even small variations in temperature Heat Pulse Duration and Temperature: Optimal transformation efficiency is observed when 100 μl of cells are heat-pulsed at 42°C for 33 seconds. Efficiency decreases sharply when cells are heat-pulsed for 35 seconds. Do not exceed 42°C. then you may follow the guide instructions.
Pls. find the attached files
http://www.yeastern.com/Products.php?pkid=8
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3860579/
http://www.bioline.org.br/pdf?ej05013
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC306974/pdf/nar00011-0160.pdf
http://www.ejbiotechnology.cl/index.php/ejbiotechnology/article/view/v8n1-8/442
http://link.springer.com/article/10.1007/BF02761755
http://link.springer.com/protocol/10.1385/0-89603-332-5:31
http://johnson.cm.utexas.edu/kjohnson/methods/Inoue_Bacterial_Transformation.php
Hoping this will be helpful
Regards
Prof. Houda Kawas
You can make competent cells using Cesium Chloride instead of Calcium chloride. That will surely help you in better transformation efficiency.
The best condition for transformation is from 37-42oC. Over than 42oC will make the bacterial die. To increase the transformation efficiency, you must improve the competent cells. In this case, I think there could be due to the competent cells or due to the big plasmids.
You shouldn't go over 42 °C for transformation. Certain proteins tend to have very sharp denaturation curves. Even small differences in temperature can have large influences on bacterial viability, and therefore transformation efficiency.
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