Tag Archives: mechanical

Finishing out with the mechanical

Currently there is a pretty robust spreadsheet and over the past few days Jake has cranked through some reps to see how the oysters that were mechanically stressed hold up. Below is how these data are integrated.

Jake_Heare_Research_Central__8_10_2015_EF1d_Mech_Stress_2_rep_qPCR_1B821B44.png

Currently the 8-10 samples (yellow) have been skipped, but we might have a look.

First up is having a look at the new HSP 70 reps. The mechanical data still needs some better resolution. Hopefully teh 8-10 samples migh shed some light.

Screenshot_8_17_15__9_53_AM_1B821ECC.png

Next up is two more reps of PGEEP4.
Looks good, and given the doubling of reps we could easily drop ‘outlier’ runs and still have triplicates, tight triplicates.

Screenshot_8_17_15__10_15_AM_1B822404.png

GRB2… now good to go, with the first pair of reps dead on.

BMP2…. could use some help from the other mechanical stress runs
Screenshot_8_17_15__11_03_AM_1B822F48.png

TLR….seemed like a relatively easy fix (besides no detection) in that just needed to correct for machine.
Screenshot_8_17_15__11_10_AM_1B8230E7.png

And the correction indicating the fact that expression was so low, only able to be detected by Opticon
Screenshot_8_17_15__11_17_AM_1B82327D.png

The 8-15 runs had minimal control and temp samples with mechanical run in dups.

Screenshot_8_17_15__11_24_AM_1B82344E.png

This needs a little carressing before integrating into data.
This should be in two columns with empty cells where no samples were run- in this order.

H_C_1
H_C_2
H_C_3
H_C_4
H_C_5
H_C_6
H_C_7
H_C_8
N_C_1
N_C_2
N_C_3
N_C_4
N_C_5
N_C_6
N_C_7
N_C_8
S_C_1
S_C_2
S_C_3
S_C_4
S_C_5
S_C_6
S_C_7
S_C_8
H_T_1
H_T_2
H_T_3
H_T_4
H_T_5
H_T_6
H_T_7
H_T_8
N_T_1
N_T_2
N_T_3
N_T_4
N_T_5
N_T_6
N_T_7
N_T_8
S_T_1
S_T_2
S_T_3
S_T_4
S_T_5
S_T_6
S_T_7
S_T_8
H_M_1
H_M_2
H_M_3
H_M_4
H_M_5
H_M_6
H_M_7
H_M_8
N_M_1
N_M_2
N_M_3
N_M_4
N_M_5
N_M_6
N_M_7
N_M_8
S_M_1
S_M_2
S_M_3
S_M_4
S_M_5
S_M_6
S_M_7
S_M_8

8-15 run update

Actin
Screenshot_8_17_15__3_55_PM_1B8273B6.png

Mechanical looks decent after correcting.

However taken together, bothersome the difference in crude expression levels.
Screenshot_8_17_15__3_57_PM_1B827441.png

Carm

Had some wet works issues

Jake_Heare_Research_Central__8_15_2015_CARM_CTM2_reps_qPCR_1B8274C8.png

H2AV
Assuming correction is correct- still a big differences in mechanincal here- could be real.

Screenshot_8_17_15__4_11_PM_1B8277A0.png

PGRP
No correction required as these were run on cfx, downside is some reps are not detected that would have been picked up with Opticon.

Do not see be shift in expression of mechanical stressed.
Screenshot_8_17_15__4_18_PM_1B827900.png

CRAF
Easy correction but skeptical of some very, very low Cts

Screenshot_8_17_15__4_25_PM_1B827AA2.png

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