[Gmod-gbrowse] [Gmod-help] GMOD for Next Generation Sequencing

Selvaraj, Balamuruganand (B) BSelvaraj at dow.com
Thu Jun 11 17:22:47 EDT 2009


Dave, Lincoln,
 
Finally it worked !
As told by Dave - SAMTools-0.1.3 only works.
I had to add -fPIC parameter with CFLAG  in Makefile of SAMTools-0.1.3.
 
-Bala
 
________________________________

From: Dave Clements, GMOD Help Desk [mailto:gmodhelp at googlemail.com] 
Sent: Thursday, June 11, 2009 4:43 PM
To: Selvaraj, Balamuruganand (B)
Cc: gmod-gbrowse at lists.sourceforge.net
Subject: Re: [Gmod-gbrowse] [Gmod-help] GMOD for Next Generation
Sequencing



	Hi Bala,
	
	I got GBrowse 2 from CPAN:
	$ sudo perl -MCPAN -e 'install Bio::Graphics::Browser'
	And I did not specify a tag on the checkout of the adaptors
(which I think means I got HEAD).
	$ cvs
-d:pserver:anonymous at gmod.cvs.sourceforge.net:/cvsroot/gmod co
gbrowse-adaptors
	
	Hope this helps,
	Dave C.
	
	On Thu, Jun 11, 2009 at 3:29 PM, Selvaraj, Balamuruganand (B)
<BSelvaraj at dow.com> wrote:
	

		Dave,
		 
		Was the Bio-SAM adaptor taken from Main Branch with CVS
tag Head.
		Thanks


________________________________

			From: Selvaraj, Balamuruganand (B)
[mailto:BSelvaraj at dow.com] 
			Sent: Thursday, June 11, 2009 2:17 PM
			To: help at gmod.org 

			Cc: gmod-gbrowse at lists.sourceforge.net
			Subject: Re: [Gmod-gbrowse] [Gmod-help] GMOD for
Next Generation Sequencing
			

			Dave,
			 
			Thanks a lot. I will try and let you know about
it.
			I was stuck with -fPIC compilation. But, I will
try with SAMTools 0.1.3.
			 
			-Bala


________________________________

				From: Dave Clements, GMOD Help Desk
[mailto:gmodhelp at googlemail.com] 
				Sent: Thursday, June 11, 2009 2:05 PM
				To: Selvaraj, Balamuruganand (B)
				Cc: gmod-gbrowse at lists.sourceforge.net
				Subject: Re: [Gmod-gbrowse] [Gmod-help]
GMOD for Next Generation Sequencing
				
				
				Hi Bala,
				
				Sorry for the slow response.  I'm
travelling all this week.
				
				Here's a long-winded explanation of what
I did to get this to work.  This may or may not apply to you as I am
running on a 32-bit OS and you are running on a 64-bit OS, and that may
be a key difference.
				
				The summary is I tried SAMtools 0.1.4,
but went back to SAMtools 0.1.3, although for different reasons then
what you are seeing.
				
				Hope this helps,
				
				Dave C.
				
				
				Grinding Details:
				
				I initially got the Bio-SamTools adaptor
to work with SAMtools 0.1.3.  I had a bit of trouble compiling SAMtools,
but none with the adaptor.  My trouble with SAMtools was:
				
				 samtools-0.1.3$ make
				 make[1]: Entering directory
`/home/clements/Documents/SAMtools/samtools-0.1.3'
				 gcc -c -g -Wall -O2 -m64  -D_IOLIB=2
-D_FILE_OFFSET_BITS=64   razip.c -o razip.o
				 In file included from
/usr/include/features.h:354,
				                from
/usr/include/stdio.h:28,
				                from razip.c:1:
				 /usr/include/gnu/stubs.h:9:27: error:
gnu/stubs-64.h: No such file or directory
				 make[1]: *** [razip.o] Error 1
				 make[1]: Leaving directory
`/home/clements/Documents/SAMtools/samtools-0.1.3'
				 make: *** [all-recur] Error 1
				
				Which seemed odd, as I have a 32-bit OS.
A SAMtools-help posting
(http://sourceforge.net/mailarchive/forum.php?thread_name=D9C3B7043F6C7C
408CC62636D8FD01BB982DCA%40ilmn-mail02.illumina.com&forum_name=samtools-
help) helped.  I dropped the -m64 from the makefile and it made fine.
				
				Bio-SamTools then also made fine.
				
				And then SAMtools 0.1.4 came out.  The
makefile that comes with 0.1.4 does not produce a libbam.a file.  The
release notes for 0.1.4 say: 
				
				Implemented the GNU building system.
However, currently the building system does not generate libbam.a. We
will improve this later. For the time being, `make -f Makefile.generic'
is preferred. 
				
				So I did that and it made, but then I
could not make the Bio-SamTools adaptor. When I got to the ./Build step
I got
				
				$ ./Build
				...
				cc -shared -L/usr/local/lib -o
blib/arch/auto/Bio/DB/Sam/Sam.so \
				 lib/Bio/DB/Sam.o \
	
-L/home/clements/Documents/SAMtools/samtools-0.1.4 \
				 -lbam -lz
	
/home/clements/Documents/SAMtools/samtools-0.1.4/libbam.a(bam_aux.o): \
				 In function `bam_aux_get_core':
	
/home/clements/Documents/SAMtools/samtools-0.1.4/bam_aux.c:22: \
				 multiple definition of
`bam_aux_get_core'
				lib/Bio/DB/Sam.o:Sam.c:(.text+0xb220):
first defined here
				collect2: ld returned 1 exit status
				error building
blib/arch/auto/Bio/DB/Sam/Sam.so from lib/Bio/DB/Sam.o \
				 at
/usr/share/perl5/ExtUtils/CBuilder/Base.pm line 212.
				
				And then, I gave up and went back to
0.1.3.  And that worked fine.  I did all this on the version of
Bio-SamTools that existed in CVS on 2009/05/21.
				
				On Thu, Jun 11, 2009 at 8:46 AM,
Selvaraj, Balamuruganand (B)<BSelvaraj at dow.com> wrote:
				> Hi,
				>  
				> I tried to recompile with -fPIC
option, still get the same error. Has anyone
				> tried to use the Bio:DB:SAM plugin
with SAMtools ?
				> Let me know which version of the
SAMTools you used ? I doubt if the latest
				> version(0.1.4) of SAMTools is having
some problem
				>  
				> Thanks
				> -Bala
				>
				> ________________________________
				> From: Selvaraj, Balamuruganand (B)
				> Sent: Wednesday, June 10, 2009 1:59 PM
				> To: 'Lincoln Stein'
				> Cc:
'gmod-gbrowse at lists.sourceforge.net'
				> Subject: RE: [Gmod-gbrowse]
[Gmod-help] GMOD for Next Generation Sequencing
				>
				> Lincoln,
				>  
				> I was trying to build the Bio/Sam
pckage obtained from the repository.
				> However, the Build.PL present in
Bio-SamTools runs without any issues
				> But, when I try ./Build I get the
below error.
				>  
				> gcc -shared -O2 -g -pipe -Wall
-Wp,-D_FORTIFY_SOURCE=2 -fexceptions
				> -fstack-protector
--param=ssp-buffer-size=4 -m64 -mtune=generic -o
				> blib/arch/auto/Bio/DB/Sam/Sam.so
lib/Bio/DB/Sam.o -L/home/samtools-0.1.4
				> -lbam -lz
				> /usr/bin/ld:
/home/n006481/samtools-0.1.4/libbam.a(bam.o): relocation
				> R_X86_64_32 against `a local symbol'
can not be used when making a shared
				> object; recompile with -fPIC
				> /home/samtools-0.1.4/libbam.a: could
not read symbols: Bad value
				> collect2: ld returned 1 exit status
				> error building
blib/arch/auto/Bio/DB/Sam/Sam.so from lib/Bio/DB/Sam.o at
				>
/usr/lib/perl5/site_perl/5.8.8/ExtUtils/CBuilder/Base.pm line 212.
				>  
				> When I debugged for the error - it
seems to be coming from $build
				> -> dispatch the last line of the Build
				> It seems to me to be PIC error. Is
there a way by which we can rectify this
				> error ?
				>  
				> -Bala
				>
				-- 
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				Learn more about GMOD at:
				 Arthropod Genomics:
http://www.k-state.edu/agc/symp2009/seminar.html
				 AGA Next Gen Seq in Non-Models:
http://www.regonline.com/Nextgeneration
				
				


	
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	-- 
	Was this helpful?  Let us know at
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	Learn more about GMOD at:
	 SMBE:
http://ccg.biology.uiowa.edu/smbe/symposia.php?action=view&sym_ID=27
	 Arthropod Genomics:
http://www.k-state.edu/agc/symp2009/seminar.html
	 AGA Next Gen Seq in Non-Models:
http://www.regonline.com/Nextgeneration
	

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