further work on pcdf6 cloning, second experiment on larvae locomotion

on Monday, December 7th, 2020 12:33 | by Sarah-Lynn Stratil

PCR pCDF6 Primer
-12.5 µl primer reverse/forward
-5 µl dNTPs
-2.5 µl Polymerase
-0.57 µl pCDF6 undigested
-50 µl Q5 buffer
-166.93 µl H2O
Agarose-gel electrophoresis
E.Z.N.A. Gel Extraction

pcr1: 153.3 ng/µl
pcr2: 115.7 ng/µl
pcr3: 174.4 ng/µl

pCDF6 NEBuilder Assembly Reaction
-pcr1: 4.85 ng -> 0.88 µl
-pcr2: 4.24 ng -> 1.5 µl
-pcr3: 4.87 ng -> 1.6 µl
-pCDF6 digested: 100 ng -> 2.8 µl
-H2O: 3.22 µl
-10 µl NEBuilder HiFi DNA Assembly Master Mix/Control
PCR
heat shock transformation of the construct into competent E.coli cells
-> plated on LB0+Amp plates
-> no colonies

control: ELAV-Gal4 x UAS-gRNA (n=23)
experimental: ELAV-Gal4 x UAS-Cas9-gRNA (n=23)


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Category: Foxp, Uncategorized

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