<div dir="ltr"><div>Hi Bashir, it looks like tNCS is correctly accounted for by Phaser; your native Patterson map has a peak at {0.157 -0.000 -0.333}, which corresponds to the translation vector between your two molecules in solution 1.</div><div><br></div><div>Your problem here is probably related to this warning:</div><div><br></div><div><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Warning: The composition you have entered gives a solvent content of 87%, which is<br>extremely unlikely. You should increase the composition.<br><br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><font face="arial,sans-serif">You don't have a complete crystal lattice in your model. Check near the top of the Phaser log file for the Matthews coefficient calculation, which will suggest the most probable multiples of your input molecular weight. You can also calculate these probabilities with the ccp4 program matthews_coeff or Bernard Rupps server at <a href="http://www.ruppweb.org/mattprob/default.html">http://www.ruppweb.org/mattprob/default.html</a>.<br><br></font></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><font face="arial,sans-serif">Best wishes<br></font></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><font face="arial,sans-serif">Kevin<br></font></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i></i></pre></div><div><div><div dir="ltr" class="gmail_signature" data-smartmail="gmail_signature"><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><div><div dir="ltr"><span><div>--<br></div><div>Kevin Jude, PhD (he/him/his)<br><div>Structural Biology Research Specialist, Garcia Lab</div><div>Howard Hughes Medical Institute</div></div><div>Stanford University School of Medicine</div><div>Beckman B177, 279 Campus Drive, Stanford CA 94305</div><div>Phone: <a href="tel:%28650%29%20723-6431" value="+16507236431" target="_blank">(650) 723-6431</a></div></span></div></div></div></div></div></div></div></div></div><br></div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Thu, Feb 25, 2021 at 12:57 PM Muhammad Bashir Khan <<a href="mailto:mbk@ualberta.ca">mbk@ualberta.ca</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex"><div dir="ltr">Dear All;<div><br></div><div><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i>I collected data set which can be easily processed in a space grouop P21 (P1211)</i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i>at approx 3.7A. I procedeed with MR using phaser in phenix which give a solution with two copies.
</i><i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">** There were 8 solutions<br><br>** Pdb and/or Mtz files have been written with results for 1 of these solutions<br><br>** Solution #1 written to PDB file: Khan_phaser.1.pdb<br>** Solution #1 written to MTZ file: Khan_phaser.1.mtz<br> Solution #1 annotation (history):<br> SOLU SET RFZ=3.4 TFZ=4.2 PAK=0 LLG=15 TFZ==5.5 RFZ=2.8 TFZ=27.6 PAK=5 LLG=1173 TFZ==36.5 LLG=1210 TFZ==36.8 PAK=6<br> LLG=1210 TFZ==36.8<br> SOLU SPAC P 1 2 1<br> SOLU 6DIM ENSE ense_1 EULER 7.6 86.4 3.3 FRAC 0.20 0.00 0.18 BFAC -1.90 #TFZ==5.5<br> SOLU 6DIM ENSE ense_1 EULER 7.6 86.4 3.2 FRAC 0.05 0.00 0.51 BFAC 1.60 #TFZ==36.8<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0444 #RMSD 1.29 #VRMS 1.64<br><br> Solution #2 annotation (history):<br> SOLU SET RFZ=3.0 TFZ=5.5 PAK=0 LLG=10 RFZ=2.7 TFZ=25.6 PAK=0 LLG=1165 TFZ==34.8 LLG=1198 TFZ==34.0 PAK=4 LLG=1198<br> TFZ==34.0<br> SOLU SPAC P 1 2 1<br> SOLU 6DIM ENSE ense_1 EULER 10.7 83.5 177.9 FRAC 0.24 -0.00 0.14 BFAC -3.15<br> SOLU 6DIM ENSE ense_1 EULER 169.3 96.4 357.9 FRAC 0.91 -0.00 0.52 BFAC 1.82 #TFZ==34.0<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0705 #RMSD 1.29 #VRMS 1.65<br><br> Solution #3 annotation (history):<br> SOLU SET RFZ=3.4 TFZ=4.1 PAK=1 LLG=-1 RFZ=2.8 TFZ=34.6 PAK=1 LLG=1146 TFZ==39.7 LLG=1185 TFZ==39.2 PAK=3 LLG=1186<br> TFZ==39.2<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 8.2 86.4 2.0 FRAC 0.01 -0.00 0.01 BFAC -1.69<br> SOLU 6DIM ENSE ense_1 EULER 8.3 86.4 2.0 FRAC -0.14 -0.00 0.34 BFAC 1.72 #TFZ==39.2<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0406 #RMSD 1.29 #VRMS 1.64<br><br> Solution #4 annotation (history):<br> SOLU SET RFZ=3.5 TFZ=5.0 PAK=6 LLG=13 RFZ=2.9 TFZ=30.5 PAK=6 LLG=1156 TFZ==34.4 LLG=1183 TFZ==33.6 PAK=7 LLG=1183<br> TFZ==33.6<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 359.5 82.0 2.7 FRAC -0.18 -0.00 0.47 BFAC -2.89<br> SOLU 6DIM ENSE ense_1 EULER 359.5 82.1 2.8 FRAC -0.02 -0.00 0.14 BFAC 2.57 #TFZ==33.6<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0529 #RMSD 1.29 #VRMS 1.64<br><br> Solution #5 annotation (history):<br> SOLU SET RFZ=3.5 TFZ=5.0 PAK=6 LLG=13 RFZ=3.0 TFZ=29.0 PAK=6 LLG=1115 TFZ==34.1 LLG=1178 TFZ==33.6 PAK=7 LLG=1178<br> TFZ==33.6<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 359.9 80.8 0.7 FRAC -0.18 -0.00 0.47 BFAC -1.57<br> SOLU 6DIM ENSE ense_1 EULER 180.1 99.2 180.7 FRAC 0.34 0.50 0.20 BFAC 2.47 #TFZ==33.6<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0405 #RMSD 1.29 #VRMS 1.64<br><br> Solution #6 annotation (history):<br> SOLU SET RFZ=3.5 TFZ=4.6 PAK=1 LLG=-0 RFZ=2.8 TFZ=36.2 PAK=1 LLG=1136 TFZ==35.0 LLG=1169 TFZ==34.6 PAK=1 LLG=1169<br> TFZ==34.6<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 359.2 83.0 1.5 FRAC 0.06 0.01 0.03 BFAC -2.58<br> SOLU 6DIM ENSE ense_1 EULER 359.2 82.9 1.4 FRAC -0.09 0.01 0.36 BFAC 2.08 #TFZ==34.6<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0869 #RMSD 1.29 #VRMS 1.66<br><br> Solution #7 annotation (history):<br> SOLU SET RFZ=3.5 TFZ=4.6 PAK=1 LLG=-0 RFZ=2.8 TFZ=35.3 PAK=1 LLG=1122 TFZ==35.4 LLG=1149 TFZ==34.5 PAK=1 LLG=1148<br> TFZ==34.5<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 359.2 82.5 2.4 FRAC 0.06 0.01 0.03 BFAC -1.93<br> SOLU 6DIM ENSE ense_1 EULER 180.8 97.5 182.4 FRAC 0.78 -0.49 0.31 BFAC 2.44 #TFZ==34.5<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0809 #RMSD 1.29 #VRMS 1.65<br><br> Solution #8 annotation (history):<br> SOLU SET RFZ=3.4 TFZ=4.1 PAK=1 LLG=-1 RFZ=2.8 TFZ=33.9 PAK=1 LLG=1083 TFZ==38.5 LLG=1146 TFZ==38.5 PAK=4 LLG=1146<br> TFZ==38.5<br> SOLU SPAC P 1 21 1<br> SOLU 6DIM ENSE ense_1 EULER 8.1 86.1 2.6 FRAC 0.01 -0.00 0.00 BFAC -1.77<br> SOLU 6DIM ENSE ense_1 EULER 8.1 86.2 2.6 FRAC 0.17 -0.00 -0.33 BFAC 1.66 #TFZ==38.5<br> SOLU ENSEMBLE ense_1 VRMS DELTA +1.0762 #RMSD 1.29 #VRMS 1.65<br><br>CPU Time: 0 days 6 hrs 35 mins 53.63 secs ( 23753.63 secs)<br>Finished: Tue Feb 16 23:27:36 2021<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i>Got the following warnings:</i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">WARNINGS<br>--------<br><br>------------------------------------------------------------------------------------------<br>Warning: Intensity moments suggest possibility of twinning. Tests based on possible twin<br>laws will be more definitive.<br>------------------------------------------------------------------------------------------<br><br>------------------------------------------------------------------------------------------<br>Warning: The composition you have entered gives a solvent content of 87%, which is<br>extremely unlikely. You should increase the composition.<i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">After running xtraige for the .mtz, I get this warning about NCS<i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Frac. coord. : 0.157 -0.000 -0.333<i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Distance to origin : 119.294<i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Height relative to origin : 33.056 %<i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">p_value(height) : 8.504e-04.<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Translational pseudo-symmetry is very likely present in these data.
Be aware that this will change the intensity statistics and may impact
subsequent analyses, and in practice may lead to higher R-factors in
refinement.<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Upon an attempt at initial refinement with different stratageis, the R-fators are stuck at<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Start R-work = 0.5066, R-free = 0.5233<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Final R-work = 0.4831, R-free = 0.4965<br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><br></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">The question is that there is something that is not being recognized
for example merohedral twinning or that the NCS/pseudosymmetry is not <span style="font-family:Arial,Helvetica,sans-serif">being addressed</span></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"> correctly. Any suggestions would be highly appreciated.</pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Regards;</pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)"><i><br></i></pre><pre style="white-space:pre-wrap;color:rgb(0,0,0)">Bashir</pre></div><div><div>-- <br></div><div dir="ltr"><div dir="ltr"><div><div dir="ltr"><div><span style="font-family:Helvetica;font-size:medium">------------------------------------------------------</span></div><span style="font-family:Helvetica;font-size:medium">Muhammad Bashir Khan, Ph.D.</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Research Associate</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Department of Biochemistry</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Medical Science Bldg.</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Lab 3-27</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">University of Alberta</span><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Edmonton AB, T6G 2H7</span><br style="font-family:Helvetica;font-size:medium"><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">Phone: </span><a value="+17804923465" style="color:rgb(17,85,204);font-family:Helvetica;font-size:medium">780-492-4577-</a><br style="font-family:Helvetica;font-size:medium"><span style="font-family:Helvetica;font-size:medium">e-mail: </span><a href="mailto:cbrooks1@ualberta.ca" style="color:rgb(17,85,204);font-family:Helvetica;font-size:medium" target="_blank">mbk@ualberta.ca</a><br></div></div></div></div></div></div>
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