Hi David,

Thank you for your explanation and suggestions! I will install Java 9 JDK 
and update the Windows PATH accordingly.

You mentioned several important settings for PeptideProphet in MSFragger 
searches. Could you help me check if this setting is correct?
[image: Snipaste_2024-05-10_21-21-43.png]

 If I want to perform TMT quantification of peptides with specific PTMs, 
which modules should I run after the main search? I assume the workflow 
would be like PeptideProphet -> PTMProphet -> ProteinProphet -> Libra.  

I'm new to TPP, though I attended your poster session previously. Could you 
recommend any detailed tutorial resources for using MSFragger in TPP?

Thank you!
Zeyu Wang 



On Friday, May 10, 2024 at 6:55:36 PM UTC-4 David Shteynberg wrote:

> Hi  Zeyu,
>
> Thanks for your question and for your interest in the TPP.
>
> Yes I use MSFragger in the TPP and I did run into this exact problem last 
> week.  It required me to install java 9 jdk that I downloaded and installed 
> from oracle.  I then had to make sure that the windows PATH pointed to the 
> correct version of java from the jdk.  Let me know if you cannot find it 
> and I would be glad to help you further.
>
> I have found that the PeptideProphet model that seems to function best for 
> MSFragger searches is the EXPECTSCORE based NONPARAM (semi-supervised) 
> model, also with the option ONEFVAL to model all the charge states with a 
> single f-value mixture model.
>
> Hope you find this useful. 
>
> Cheers!
> -David
>
> On Fri, May 10, 2024 at 3:05 PM Zeyu Wang <[email protected]> wrote:
>
>> Hi,
>>
>> Do you use MSFragger in TPP? I downloaded MSFragger 4.0 and renamed it, 
>> then placed it in this path: C:/TPP/bin/msfragger/msfragger.jar. However, 
>> when I search my files, it seems to report an error related to the JRE 
>> version. But MSFragger desktop GUI runs fine:
>>
>> Error: A JNI error has occurred, please check your installation and try 
>> again.
>> Exception in thread "main" java.lang.UnsupportedClassVersionError: 
>> edu/umich/andykong/msfragger/MSFragger has been compiled by a more recent 
>> version of the Java Runtime (class file version 53.0), this version of the 
>> Java Runtime only recognizes class file versions up to 52.0.
>>     at java.lang.ClassLoader.defineClass1(Native Method)
>>     at java.lang.ClassLoader.defineClass(Unknown Source)
>>     at java.security.SecureClassLoader.defineClass(Unknown Source)
>>     at java.net.URLClassLoader.defineClass(Unknown Source)
>>     at java.net.URLClassLoader.access$100(Unknown Source)
>>     at java.net.URLClassLoader$1.run(Unknown Source)
>>     at java.net.URLClassLoader$1.run(Unknown Source)
>>     at java.security.AccessController.doPrivileged(Native Method)
>>     at java.net.URLClassLoader.findClass(Unknown Source)
>>     at java.lang.ClassLoader.loadClass(Unknown Source)
>>     at sun.misc.Launcher$AppClassLoader.loadClass(Unknown Source)
>>     at java.lang.ClassLoader.loadClass(Unknown Source)
>>     at sun.launcher.LauncherHelper.checkAndLoadMain(Unknown Source)
>>
>>
>> Thank you!
>>
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