This masterclass was authored by Giovanni Bussi on February 15, 2021.
In this Masterclass, we will discuss how to perform and analyze umbrella sampling simulations. We will learn how to introduce a bias potential with PLUMED, how to compute the free energy landscapes, and how to reweight the resulting ensembles. We will also learn how to compute statistical errors on the computed quantities.
Once you have completed this Masterclass you will be able to:
- Use PLUMED to run simulations using static bias potentials with different functional forms.
- Use WHAM to combine multiple simulations performed with different bias potentials.
- Reweight the resulting ensembles so as to obtain the free-energy profile as a function of a different variable.
- Calculate error bars on free energies and populations.
If you have not yet set up PLUMED, you can find information about installing it here.
Once you have installed PLUMED, you will need to install GROMACS as well. In particular, you will need a special version of GROMACS that has been patched with PLUMED. You can obtain it by using conda with the following command
conda install --strict-channel-priority -c plumed/label/masterclass -c conda-forge gromacs
The --strict-channel-priority
is necessary as it prevents your conda install from downloading packages from the bioconda
channel. bioconda
contains a version of GROMACS that is not patched with PLUMED and would not work here.
On Linux, the command above should install the following packages:
gromacs plumed/label/masterclass/linux-64::gromacs-2019.6-h3fd9d12_0
libclang conda-forge/linux-64::libclang-11.0.1-default_ha53f305_1
libevent conda-forge/linux-64::libevent-2.1.10-hcdb4288_3
libhwloc conda-forge/linux-64::libhwloc-1.11.13-h3c4fd83_0
libllvm11 conda-forge/linux-64::libllvm11-11.0.1-hf817b99_0
libpq conda-forge/linux-64::libpq-12.3-h255efa7_3
[ etc ... ]
The exact versions might be different. Notice, however, that GROMACS comes from the plumed/label/masterclass
channel, whereas the required libraries come from the conda-forge
channel. To be sure the installed GROMACS is patched with PLUMED, try the following shell command:
gmx mdrun -h 2> /dev/null | grep -q plumed && echo ok
It should print ok
.
Please ensure that you have setup PLUMED and GROMACS on your machine before starting the exercises. Also notice that in order to obtain a good performance it is better to compile GROMACS from source on the machine where you are running your simulations. You can find out from the PLUMED documention how to patch GROMACS with PLUMED so as to be able to install it from source. For this tutorial, the conda precompiled binaries will be sufficient.
The data needed to execute the exercises of this Masterclass can be found on GitHub. You can clone this repository locally on your machine using the following command:
git clone https://github.com/plumed/masterclass-21-3.git
The files you need for the exercise are in the folder called data
. You will find the following files in that folder:
topolA.tpr
: an input file that can be used to run a GROMACS simulation of alanine dipeptide starting from one of the two main free-energy minimatopolB.tpr
: same astopolA.tpr
, but starting from the other minimum.wham.py
: a python script that can be used to perform binless WHAM analysisreference.pdb
: a pdb file of alanine dipeptide (the system under study) that can be used as input for your MOLINFO commands
Notice that PLUMED input files have not been provided in the GitHub repository. You must prepare these input files yourself using the templates below.
We would recommend that you run each exercise in separate sub-directories inside the root directory masterclass-21-3
.
All the exercises were tested with PLUMED version 2.7.0 and GROMACS 2019.6
Throughout this tutorial we will run simulations of alanine dipeptide in vacuum using GROMACS and PLUMED. Whereas this system is too simple to be considered a proper benchmark for enhanced sampling methods, it is complex enough to be used when learning about them. Some of the commands below are specific for GROMACS, but all the PLUMED input files are compatible with other MD engines as well.
In other masterclasses you learned how to analyze trajectories a posteriori. One of the nice features of PLUMED is that the very same analysis can be done on the fly. In other words, you might compute your collective variables while GROMACS is running instead of waiting for the simulation to end. This can could be convenient if you want to run a large number of simulations, you already know what to compute, and you do not want to use too much disk space.
To run a simulation with GROMACS you have to type this command in the shell:
gmx mdrun -plumed plumed.dat -s topolA.tpr -nsteps 200000 -x traj_unbiased.xtc
Notice that the file topolA.tpr
contains all the relevant information (simulation parameters, initial conditions, etc.). In this tutorial we will just need to play with the number of steps (200000 in the example above) and we will tune the name of the trajectory saved by GROMACS (traj_unbiased.xtc in the example above).
Also consider that GROMACS and PLUMED will never delete your files. They will take backups when you try to overwrite them. GROMCAS backup files start with #
, whereas PLUMED backup files start with bck.
. Both GROMACS and PLUMED will complain if you try to create too many backups of the same file. It is thus recommended to regularly clean your directory with a command such as rm -f \#* bck.*
The command above will only succeed if a file named plumed.dat
exists in the current directory. You should already know to create such file from having completed other (more basic) tutorials. Those other tutorials should also have taught you how to compute histograms. You should be thus able to complete the template below and put it in a file named plumed.dat
.
#SOLUTIONFILE=work/plumed_ex1.dat
# vim:ft=plumed
MOLINFO STRUCTURE=reference.pdb
phi: TORSION ATOMS=__FILL__ # use MOLINFO shortcuts to identify the phi angle of the second residue
psi: TORSION ATOMS=__FILL__ # use MOLINFO shortcuts to identify the psi angle of the second residue
# use the command below to compute the histogram of phi
# we use a smooth kernel to produce a nicer graph here
# notice that when asking for numbers PLUMED is happy to accept strings such as "pi" meaning 3.14...
# also arithmetics is allowed (e.g. 2*pi could be used if necessary)
hhphi: HISTOGRAM ARG=__FILL__ STRIDE=100 GRID_MIN=-pi GRID_MAX=__FILL__ GRID_BIN=600 BANDWIDTH=0.05
ffphi: CONVERT_TO_FES GRID=hhphi # no need to set TEMP here, PLUMED will obtain it from GROMACS
DUMPGRID GRID=__FILL__ FILE=fes_phi.dat STRIDE=200000 # stride is needed here since PLUMED does not know when the simulation is over
# now add three more lines to compute and dump the free energy as a function of **psi** on a file names fes_psi.dat
__FILL__
PRINT __FILL__ # use this command to write phi and psi on a file named colvar.dat, every 100 steps
You can then monitor what happened during the simulation using the python script below.
import plumed
import matplotlib.pyplot as plt
import numpy as np
# plot the time series of phi and psi
colvar=plumed.read_as_pandas("colvar.dat")
plt.plot(colvar.time,colvar.phi,"x",label="phi")
plt.plot(colvar.time,colvar.psi,"x",label="psi")
plt.xlabel("time")
plt.ylabel("$\phi$")
plt.legend()
plt.show()
# scatter plot with phi and psi
plt.plot(colvar.phi,colvar.psi,"x")
plt.xlabel("$\phi$")
plt.ylabel("$\psi$")
plt.xlim((-np.pi,np.pi))
plt.ylim((-np.pi,np.pi))
plt.show()
# FES as a function of phi
# we remove infinite and nans here
fes_phi=plumed.read_as_pandas("fes_phi").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phi.phi,fes_phi.ffphi)
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\phi$")
plt.ylabel("$F(\phi)$")
plt.show()
# FES as a function of psi
# we remove infinite and nans here
fes_psi=plumed.read_as_pandas("fes_psi").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psi.psi,fes_psi.ffpsi)
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\psi$")
plt.ylabel("$F(\psi)$")
plt.show()
In this manner you will be able to see (a) the time series of
We are now ready to use PLUMED to perform the task it was originally designed for: biasing a simulation on the fly. We will first try to add simple bias potentials that change the balance between the two minima we have obtained in the previous exercise. Biasing is not particularly useful here since both minima were sampled anyways. However, it is instructive since we will be able to test the types
of analysis we did in masterclass-21-2. The two minima observed in the previous exercise are located at -A*sin(x+2)
, with
We can try this type of calculation with A=10 by using an input file like the following one:
#SOLUTIONFILE=work/plumed_ex2.dat
# vim:ft=plumed
__FILL__ # compute phi and psi here, as in the previous input file
# fill in with the required function (i.e. -10*sin(phi+2)):
f: CUSTOM ARG=phi FUNC=__FILL__ PERIODIC=NO
# this command allows to add a bias potential equal to f
BIASVALUE ARG=f
# here you can paste the same HISTOGRAM/CONVERT_TO_FES/DUMPGRID commands that you used in
# the previous exercise. let's just write the results on different files
hhphi: __FILL__
ffphi: __FILL__
DUMPGRID FILE=fes_phi_biased1.dat __FILL__
hhpsi: __FILL__
ffpsi: __FILL__
DUMPGRID FILE=fes_psi_biased1.dat __FILL__
lw: REWEIGHT_BIAS
# and here we do the same again, but this time using LOGWEIGHTS.
# these free energies will be printed on files fes_phi_biased1r.dat and
# fes_psi_biased1r.dat and will be reweighted so as to be unbiased
hhphir: HISTOGRAM __FILL__ LOGWEIGHTS=lw
ffphir: __FILL__
DUMPGRID FILE=fes_phi_biased1r.dat __FILL__
hhpsir: HISTOGRAM __FILL__ LOGWEIGHTS=lw
ffpsir: __FILL__
DUMPGRID FILE=fes_psi_biased1r.dat __FILL__
PRINT __FILL__ # monitor what's happening, as before, writing on file plumed_colvar1.dat
Call this file plumed_biased1.dat
and run the simulation using this command:
gmx mdrun -plumed plumed_biased1.dat -s topolA.tpr -nsteps 200000 -x traj_comp_biased1.xtc
Notice that we are storing the trajectory on a separate file. We will need both the unbiased and the biased trajectory later.
You can then monitor what happened during the simulation using these commands
colvar=plumed.read_as_pandas("colvar_biased1.dat")
plt.plot(colvar.time,colvar.phi,"x",label="phi")
plt.plot(colvar.time,colvar.psi,"x",label="psi")
plt.xlabel("time")
plt.ylabel("$\phi$")
plt.legend()
plt.show()
plt.plot(colvar.phi,colvar.psi,"x")
plt.xlabel("$\phi$")
plt.ylabel("$\psi$")
plt.xlim((-np.pi,np.pi))
plt.ylim((-np.pi,np.pi))
plt.show()
fes_phi=plumed.read_as_pandas("fes_phi.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phi.phi,fes_phi.ffphi,label="original")
fes_phib=plumed.read_as_pandas("fes_phi_biased1.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phib.phi,fes_phib.ffphi,label="biased")
fes_phir=plumed.read_as_pandas("fes_phi_biased1r.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phir.phi,fes_phir.ffphir,label="reweighted")
plt.legend()
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\phi$")
plt.ylabel("$F(\phi)$")
plt.show()
fes_psi=plumed.read_as_pandas("fes_psi.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psi.psi,fes_psi.ffpsi,label="original")
fes_psib=plumed.read_as_pandas("fes_psi_biased1.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psib.psi,fes_psib.ffpsi,label="biased")
fes_psir=plumed.read_as_pandas("fes_psi_biased1r.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psir.psi,fes_psir.ffpsir,label="reweighted")
plt.legend()
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\psi$")
plt.ylabel("$F(\psi)$")
plt.show()
The free-energy plots here will include three lines: (a) the original one (as obtained from the previous exercise), (b) the one sampled in this second simulation, where the minimum at higher phi appears more stable, and (c) the reweighted free-energy from the second simulation. This final free energy surface should look closer to the original one. Notice that the relationship between plots (b) and (c) is straightforward when the analyzed variable is phi: the third line could have been obtained by just adding
We will now run a new simulation that is even more biased by choosing the prefactor A=20. This will further favour the minimum at higher values of phi. Please prepare a plumed_biased2.dat
input file that is identical to plumed_biased1.dat
except that:
- it applies a bias with -20*sin(phi+2)
- all the written files are named with a
2
suffix (e.g.fes_phi_biased2.dat
).
Then run the new simulation using the following command:
gmx mdrun -plumed plumed_biased2.dat -s topolA.tpr -nsteps 200000 -x traj_comp_biased2.xtc
You can analyze this new simulation as you did the previous one. You should notice that the peak at higher
We will now use WHAM to combine the three simulations we have done so far, namely:
- unbiased (
traj_comp_unbiased.xtc
), - biased (
traj_comp_biased1.xtc
), and - more biased (
traj_comp_biased2.xtc
).
The first thing we have to do is to concatenate the three files usinig:
gmx trjcat -cat -f traj_comp_unbiased.xtc traj_comp_biased1.xtc traj_comp_biased2.xtc -o traj_comp_cat.xtc
Notice that this new trajectory contains all the simulated frames, and we do not explicitly track which of the simulation each frame originated from.
We then have to compute the bias that would have been felt in each of the three runs above by each of the frames in the concatenated trajectory. We could do this reusing the three plumed input files we used above, but it is perhaps clearer to do it with a single input file in this case. You can use an input like this one:
#SOLUTIONFILE=work/plumed_ex3.dat
# vim:ft=plumed
MOLINFO STRUCTURE=reference.pdb
phi: TORSION __FILL__
psi: TORSION __FILL__
# here we list the bias potentials that we used in the three simulations we want to combine
b0: CUSTOM ARG=phi FUNC=0.0 PERIODIC=NO
b1: CUSTOM ARG=phi FUNC=__FILL__ PERIODIC=NO # fill here with the bias you used in the first biased simulation
b2: CUSTOM ARG=phi FUNC=__FILL__ PERIODIC=NO # fill here with the bias you used in the second biased simulation
PRINT ARG=phi,psi,b0,b1,b2 FILE=biases.dat
If you call your input file plumed_wham.dat
you can then use the following command
plumed driver --plumed plumed_wham.dat --ixtc traj_comp_cat.xtc
We are now ready to use binless WHAM to compute the weights associated to the concatenated trajectory. This can be done using the following script
import wham
# this is to check that you actually imported the module located in this directory
# it should print /current/directory/wham.py
print(wham.__file__)
bias=plumed.read_as_pandas("biases.dat")
kBT=300*8.314462618*0.001 # use kJ/mol here
w=wham.wham(np.stack((bias.b0,bias.b1,bias.b2)).T,T=kBT)
print(w)
# you can then add a new column to the bias dataframe:
print(bias)
bias["logweights"]=w["logW"]
print(bias)
# and write it on disk
plumed.write_pandas(bias,"bias_wham.dat")
The procedure above computes the weights that can be associated to the concatenated trajectory so as to recover an unbiased distribution. Notice that, although we did not explicitly enforce it,
the weights only depend on the value of
plt.plot(bias.phi,bias.logweights,"x")
plt.show()
plt.plot(bias.psi,bias.logweights,"x")
plt.show()
Also notice the form of the weights as a function of
We can then read the obtained weights and use them in an analysis similar to the ones we have done above
#SOLUTIONFILE=work/plumed_ex4.dat
# vim:ft=plumed
phi: READ FILE=bias_wham.dat VALUES=__FILL__
psi: READ FILE=bias_wham.dat VALUES=__FILL__
lw: READ FILE=bias_wham.dat VALUES=__FILL__
hhphi: HISTOGRAM ARG=phi GRID_MIN=-pi GRID_MAX=pi GRID_BIN=600 BANDWIDTH=0.05
ffphi: CONVERT_TO_FES GRID=hhphi
DUMPGRID GRID=ffphi FILE=fes_phi_cat.dat
hhpsi: HISTOGRAM ARG=psi GRID_MIN=-pi GRID_MAX=pi GRID_BIN=600 BANDWIDTH=0.05
ffpsi: CONVERT_TO_FES GRID=hhpsi
DUMPGRID GRID=ffpsi FILE=fes_psi_cat.dat
# we use a smooth kernel to produce a nicer graph here
hhphir: HISTOGRAM ARG=phi GRID_MIN=-pi GRID_MAX=pi GRID_BIN=600 BANDWIDTH=0.05 LOGWEIGHTS=lw
ffphir: CONVERT_TO_FES GRID=hhphir
DUMPGRID GRID=ffphir FILE=fes_phi_catr.dat
hhpsir: HISTOGRAM ARG=psi GRID_MIN=-pi GRID_MAX=pi GRID_BIN=600 BANDWIDTH=0.05 LOGWEIGHTS=lw
ffpsir: CONVERT_TO_FES GRID=hhpsir
DUMPGRID GRID=ffpsir FILE=fes_psi_catr.dat
Use the template above to produce a file names plumed_wham.dat
and run the following command:
plumed driver --noatoms --plumed plumed_wham.dat --kt 2.4943387854
You can now show the result by using the following script
colvar=plumed.read_as_pandas("bias_wham.dat")
plt.plot(colvar.time,colvar.phi,"x",label="phi")
plt.plot(colvar.time,colvar.psi,"x",label="psi")
plt.xlabel("time")
plt.ylabel("$\phi$")
plt.legend()
plt.show()
plt.plot(colvar.phi,colvar.psi,"x")
plt.xlabel("$\phi$")
plt.ylabel("$\psi$")
plt.xlim((-np.pi,np.pi))
plt.ylim((-np.pi,np.pi))
plt.show()
fes_phi=plumed.read_as_pandas("fes_phi.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phi.phi,fes_phi.ffphi,label="original")
fes_phib=plumed.read_as_pandas("fes_phi_cat.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phib.phi,fes_phib.ffphi,label="biased")
fes_phir=plumed.read_as_pandas("fes_phi_catr.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_phir.phi,fes_phir.ffphir,label="reweighted")
plt.legend()
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\phi$")
plt.ylabel("$F(\phi)$")
plt.show()
fes_psi=plumed.read_as_pandas("fes_psi.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psi.psi,fes_psi.ffpsi,label="original")
fes_psib=plumed.read_as_pandas("fes_psi_cat.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psib.psi,fes_psib.ffpsi,label="biased")
fes_psir=plumed.read_as_pandas("fes_psi_catr.dat").replace([np.inf, -np.inf], np.nan).dropna()
plt.plot(fes_psir.psi,fes_psir.ffpsir,label="reweighted")
plt.legend()
plt.xlim((-np.pi,np.pi))
plt.xlabel("$\psi$")
plt.ylabel("$F(\psi)$")
plt.show()
When you look at the first graph (i.e., the concatenated time series) you should notice that as you proceed towards the end the frames are coming from the most biased simulation, and thus will have a systematically larger value of
So far we only considered the fast transition between the two minima at negative values of
In the context of the static potentials that we are using here, this is a very difficult approach. In the tutorial lugano-2 you can find an attempt to derive a potential that cancels the large barrier between the states. When this bias is used the number of transitions between the important metastable states increases dramatically. In this class we will take a more pragmatic approach and directly introduce the method that is commonly used in these situations, namely "multiple windows umbrella sampling".
To do so we will have to run a number of separate simulations, each of them with a bias potential designed to maintain the
We will use 32 simulations, with RESTRAINT
potentials centered at uniformly spaced values of
import numpy as np
at=np.linspace(-np.pi,np.pi,32,endpoint=False)
print(at)
Now you should write a python script that generate 32 plumed input files (you can call them plumed_0.dat
, plumed_1.dat
, etc)
that look like this one
#SOLUTIONFILE=work/plumed_ex5.dat
# vim:ft=plumed
MOLINFO STRUCTURE=reference.pdb
phi: TORSION ATOMS=__FILL__
psi: TORSION ATOMS=__FILL__
bb: RESTRAINT ARG=phi KAPPA=200.0 AT=__FILL__
PRINT ARG=phi,psi,bb.bias FILE=__FILE__ STRIDE=100
# make sure that each simulation writes on a different file
# e.g. colvar_multi_0.dat, colvar_multi_1.dat, ...
Now run the 32 simulations with commands like these ones
gmx mdrun -plumed plumed_0.dat -s topolA.tpr -nsteps 200000 -x traj_comp_0.xtc
gmx mdrun -plumed plumed_1.dat -s topolA.tpr -nsteps 200000 -x traj_comp_1.xtc
# etc.
As we did before, you should now concatenate the resulting trajectories
gmx trjcat -cat -f traj_comp_[0-9].xtc traj_comp_[0-9][0-9].xtc -o traj_multi_cat.xtc
and analyze them with plumed driver:
plumed driver --plumed plumed_0.dat --ixtc traj_multi_cat.xtc --trajectory-stride 100
plumed driver --plumed plumed_1.dat --ixtc traj_multi_cat.xtc --trajectory-stride 100
# etc.
Notice the --trajectory-stride
option: we are telling the driver that the trajectory was saved every 100 frames. In this manner, we will be able to recycle the same plumed input file that we used while running the simulations. This approach is more robust than the one we used above, where a single plumed file was used to reproduce all the biases, and is thus recommended in production cases.
Also notice that it is common practice to ignore the initial part of each simulation, since it typically contains a transient that would make the final result less robust. For simplicitly, we are not doing it here. Finally, notice that in the last exercise we wrote a new input file just for writing the three biases. Here instea we recycled the input files used to run the 32 simulations.
We can now read the produced files and process them with our WHAM script
col=[]
for i in range(32):
col.append(plumed.read_as_pandas("colvar_multi_" + str(i)+".dat"))
# notice that this is the concatenation of 32 trajectories with 2001 frames each
plt.plot(col[i].phi[2001*i:2001*(i+1)],col[i].psi[2001*i:2001*(i+1)],"x")
plt.xlabel("$\phi$")
plt.ylabel("$\psi$")
plt.show()
# in this graph you can appreciate which region was sampled by each simulation
bias=np.zeros((len(col[0]["bb.bias"]),32))
for i in range(32):
bias[:,i]=col[i]["bb.bias"][-len(bias):]
w=wham.wham(bias,T=kBT)
plt.plot(w["logW"])
plt.show()
colvar=col[0]
colvar["logweights"]=w["logW"]
plumed.write_pandas(colvar,"bias_multi.dat")
Now you can do something similar to what we did before, namely:
- write a file to be read by PLUMED.
- process it with plumed driver so as to compute the free energy with respect to
$\phi$ and$\psi$ - plot the free energy as a function of
$\phi$ and$\psi$
How do the resulting free-energy profiles compares with the one we obtained before?
Now that you have a simulation that sampled all the relevant metastable states, you can analyze them in many different ways. In particular, you can compute the average of any quantity just using the weights that we obtained. This can be done directly with PLUMED, as we have seen in other masterclasses, by using the AVERAGE command. However, we will use Python to compute averages here as this will make it easier to compute their statistical errors with bootstrap.
To complete this exercise you should compute the average value and the statistical error of the following quantities:
- The population of the metastable state B defined as
$0<\phi<2$ . Notice that the exact definition is not very important if the minimum is clearly delimited by high free-energy barriers (i.e., low probability regions). - Free-energy difference between the state B defined as
$0<\phi<2$ and the state A defined as$-pi<\phi<0$ . Notice that the free-energy difference is defined as$-k_B T \log\left( \frac{P_B}{P_A}\right)$ , where$P_B$ is the population of state B and$P_A$ the population of state A. - Average value of
$\phi$ in state A and in state B. Warning: be careful computing the average of an angle? Remember this quantity is periodic and that must be taken into account.
For each of these quantities, you should also compute the standard error. It is here recommended to compute the error using boostrap. You can do so by adjusting the following script
# number of blocks
NB=10
# we reshape the bias so that it appears as NB times frames of each traj times number of biases
bb=bias.reshape((32,-1,32))[:,-2000:,:].reshape((32,NB,2000//NB,32))
# we reshape the trajectory so that it appears as NB times frames of each traj times number of biases
cc=np.array(col[0].phi).reshape((32,-1))[:,-2000:].reshape((32,NB,2000//NB))
# we first analyse the complete trajectory:
tr=cc.flatten()
is_in_B=np.int_(np.logical_and(tr>0,tr<2))
w0=wham.wham(bb.reshape((-1,32)),T=kBT)
print("population:",np.average(is_in_B,weights=np.exp(w0["logW"])))
pop=[]
for i in range(200):
# we then analyze the bootstrapped trajectories
print(i)
c=np.random.choice(NB,NB)
w=wham.wham(bb[:,c,:,:].reshape((-1,32)),T=kBT)
tr=cc[:,c,:].flatten()
is_in_B=np.int_(np.logical_and(tr>0,tr<2))
pop.append(np.average(is_in_B,weights=np.exp(w["logW"])))
Repeat the second to last exercise above using topolB.tpr
as an input file. This will make the simulations start from the minimum in B. Plot the free energy as a function of