Cloning of Meara RNA binding proteins

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Initial cloning of Meara RNA binding proteins. Selection: ago a, ago b, ago c, boule, bru a, bru b, gus a, mago, mex3, orb2, stau.


Only 3 genes had visible bands: boule, gus a, and stau:

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Ligated these overnight.


Transformation and plated the genes.


Colony PCR with 6 colonies from each:

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Put colonies to grow.

boule c1 BV226 BV229
boule c2 BV227 BV230
boule c3 BV228 BV231
gus a c1 BV229 BV226
gus a c2 BV230 BV227
gus a c3 BV231 BV228
stau c1 BV232
stau c2 BV233
stau c3 BV234


Extracted plasmid and did sequencing PCR for all using T7 primer. Dropped tubes into sequencing facility.


Checking sequences and they look good, but there is a problem! Minipreps BV226, 227, 228 aligned with gus a and not with boule!!! Check why…

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