ESO wikiadmin: Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=InFusion Kit Becton-Dickinson
Aqua bidest
Linearised vector (pDNR-Dual), 100ng/1µl (~4900bp)
Fragment (10-50ng/µl)
-> Subsequent method: Transformation}} == Procedure == * fragment concentration (fragment : pDNR-Dual  2:1 ratio [fmol]) * solution of a dry-down reaction in 102 µl aqua bidest * add 14 µl 10x In-Fusion-buffer and 2 µl pDNR-Dual (200 ng) * disperse to 15 tubes (8 µl per tube) and add 0.5 -1 µl of..."

2025-05-30T14:46:24Z

Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=InFusion Kit Becton-Dickinson Aqua bidest Linearised vector (pDNR-Dual), 100ng/1µl (~4900bp) Fragment (10-50ng/µl) -> Subsequent method: Transformation}} == Procedure == * fragment concentration (fragment : pDNR-Dual  2:1 ratio [fmol]) * solution of a dry-down reaction in 102 µl aqua bidest * add 14 µl 10x In-Fusion-buffer and 2 µl pDNR-Dual (200 ng) * disperse to 15 tubes (8 µl per tube) and add 0.5 -1 µl of..."

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{{Protokoll-Mix|titel=Materials|Inhalt=InFusion Kit Becton-Dickinson Aqua bidest Linearised vector (pDNR-Dual), 100ng/1µl (~4900bp) Fragment (10-50ng/µl) -> Subsequent method: Transformation}}

== Procedure ==

* fragment concentration (fragment : pDNR-Dual  2:1 ratio [fmol])
* solution of a dry-down reaction in 102 µl aqua bidest
* add 14 µl 10x In-Fusion-buffer and 2 µl pDNR-Dual (200 ng)
* disperse to 15 tubes (8 µl per tube) and add 0.5 -1 µl of the fragment (~ 2:1 ratio with regard to vector)
* fill up to 10 µl with aqua bidest and incubate for 30 min at room temperature
* put tubes on ice to stop the reaction
* transformation in ''E.coli'' nova blue and plating on LB-amp100

InFusion-cloning (Fa. Clontech) - Revision history