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.

10/03/13

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

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Ligated these overnight.

11/03/13

Transformation and plated the genes.

12/03/13

Colony PCR with 6 colonies from each:

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14/03/13

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

15/03/13

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

26/03/13

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…

Screen Shot 2013-03-26 at 10.22.12 AM