Many suppliers and users do not recommend the fast transformation protocol (Section 2.4.3) to be used to transform ligations. Why do you recommend this protocol for cloning into pSpark DNA cloning systems?
A9. pSpark DNA cloning systems are by far the most efficient cloning kits developed to date and in fact this supplied fast transformation protocol has been developed and tested to be used with pSpark DNA cloning systems. Using this fast protocol of only 5 minutes transformation, cells with a transformation efficiency of 4×107 cfu/µg and the supplied control insert, about 500-1000 positive white colonies are obtained instead of over 2500 positve colonies when using the standard protocol. If you feel uncomfortable with this protocol, use the standard protocol but as a suggestion to save your precious time we recommend to test this protocol. We do not know, however, if for example, method to prepare competent cells, E. coli strain used, temperature inside air flow cabinet, transformation protocol used or plating method have influence in outcome. To date we have no customer who have tested this fast transformation protocol for cloning into pSpark with unsatisfactory results. If you test th
