Editing FLP Recombinase mRNA (section)

== IRES-Flpo mRNA from plasmid&nbsp; ==

Product Cat #:&nbsp;EG-1045

Plasmid:&nbsp;&nbsp;138 pMiniT2-T7-IRES-Flpo

<br/>Digest plasmid with EcoR1, purify 1933&nbsp; bp fragment<br/>PCR using T7 and IVT-Flpo-R<br/>purify with column, elute with RNA-free water.

Use 1 ug as template for mRNA

T7 RNA Polymerase:

Storage Buffer:&nbsp;<br/>20 mM Tris-HC1 (pH 7.5)&nbsp;<br/>0.1 M NaCl&nbsp;<br/>0.1 mM EDTA&nbsp;<br/>1 mM DTT&nbsp;<br/>50% (v/v) glycerol&nbsp;<br/>0.01% (w/v) Triton® X-100&nbsp;&nbsp;

5X T3/T7 RNA polymerase&nbsp;Buffer:<br/>0.2 M Tris-HCl (pH 8.0)<br/>40 mM MgCl2<br/>10 mM spermidine-(HCl)3<br/>125 mM NaCl



1X Buffer:&nbsp;40 mM Tris-HCl (pH 8.0), 25 mM NaCl, 8 mM MgCl2 , 2 mM spermidine-(HCl)3 , 5 mM DTT

&nbsp;0.4 mM ATP, 0.4 mM CTP, 0.4 mM GTP, 0.4 mM UTP, in 50 µl for 10 min. at 37°C.