[nilshomer]

Hi BENN

Thank you for your script. I tried your SOLiD2Std.pl with my following data like this:

@exa1
T1011122220100230032132.2111111002.1
+
!)+%.*%*+2'0%%%-%+%*5'%!%9+'%+<+0%!%
@exa2
T0101233211103200232333.2111211002.1
+
!,.+'+')'390%%%%%%%'%%%!-<++++<99%!%
@exa3
T0312202213101213131111.1110131102.1
+
!93<*/18+%:9%+075*%:;+6!3<26%/<%-%!%


and the result is like this:

@exa1
GGTGTCTCTTGGGATTTAGTAGNNNNNNNNNNNNN
+
)+%.*%*+2'0%%%-%+%*5'%!%9+'%+<+0%!%
@exa2
TGGTCGCTGTGGCTTTCGATATNNNNNNNNNNNNN
+
,.+'+')'390%%%%%%%'%%%!-<++++<99%!%
@exa3
TACTCCTCATGGTCATGCACACNNNNNNNNNNNNN
+
93<*/18+%:9%+075*%:;+6!3<26%/<%-%!%

It seems that all letters will be converted into "N" from the first dot "."
Is that all right?

Thank you.

Without aligning (i.e. knowing the decoded DNA seqence), a missing base will not allow for a deterministic decoding (there are actually four possible sequences after a missing base).

[lix]

Hi BENN

Thank you for your script. I tried your SOLiD2Std.pl with my following data like this:

@exa1
T1011122220100230032132.2111111002.1
+
!)+%.*%*+2'0%%%-%+%*5'%!%9+'%+<+0%!%
@exa2
T0101233211103200232333.2111211002.1
+
!,.+'+')'390%%%%%%%'%%%!-<++++<99%!%
@exa3
T0312202213101213131111.1110131102.1
+
!93<*/18+%:9%+075*%:;+6!3<26%/<%-%!%


and the result is like this:

@exa1
GGTGTCTCTTGGGATTTAGTAGNNNNNNNNNNNNN
+
)+%.*%*+2'0%%%-%+%*5'%!%9+'%+<+0%!%
@exa2
TGGTCGCTGTGGCTTTCGATATNNNNNNNNNNNNN
+
,.+'+')'390%%%%%%%'%%%!-<++++<99%!%
@exa3
TACTCCTCATGGTCATGCACACNNNNNNNNNNNNN
+
93<*/18+%:9%+075*%:;+6!3<26%/<%-%!%

It seems that all letters will be converted into "N" from the first dot "."
Is that all right?

Thank you.

[BENM]

Hi BENM

I edited your SOLid2Std.pl script to include some extra colorspace mapping code that was not considered originally.

I wanted to include the following basespace mapping:
Basically any base(ATCG) that includes a '4' '5' or '.' is 'N'. 'N' to 'N' transition is also represented by diferent color space numbers (0,1,2,3,6,'.').

A4:N 
A.: N
A5:N
C4:N
C.: N
C5:N
G4:N
G.: N
G5:N
T4: N
T.:  N
T5: N
N5: A C T or G
N.:  N
N6: N
N0: N
N1: N
N2: N
N3: N

I am very naive in perl but I tried to change your script to include this conversion.
I edited the following part:

# SOLiD color code
my @code = ([0,1,2,3,'.',4,5],[1,0,3,2,'.',4,5],[2,3,0,1,'.',4,5],[3,2,1,0,'.',4,5],[5,5,5,5,'.',6,0],[5,5,5,5,1,2,3],[5,5,5,5,1,2,3]);
my @bases = qw(A C G T N N N);
my %decode = ();
foreach my $i(0..7)
{
	foreach my $j(0..7)
	{
		$decode{$code[$i]->[$j]} -> {$bases[$i]} = $bases[$j];
	}
}

It works!
However there is an error message when I run the script, although the error does not prevent it from working, which is good.
perl gives me the following error message:

Use of uninitialized value in hash element at SOLid2Std.pl line 49.
Use of uninitialized value within @bases in hash element at SOLid2Std.pl line 49
.

What am I doing wrong?
line 49 is:

$decode{$code[$i]->[$j]} -> {$bases[$i]} = $bases[$j];

Thank You

Hi inesdesantiago

Thank you for your opinions. In color space if one color space can't be recognized by SOLiD™ System, it will cause the rear bases uncertain too. So, the reads will decode "N" instead of other base in conveting color space to nucleic acid base. For expample:

@example1
G2203012023131303312303100
+
!611%%(-+%*.&*.,&2,,'%()31
@example2
G220301.023131303312303100
+
!611%%(-+%*.&*.,&2,,'%()31
@example3
G2203012023141303312303100
+
!611%%(-+%*.&*.,&2,,'%()31
@example4
G2203012023151303512303100
+
!611%%(-+%*.&*.,&2,,'%()31

There is a dot in expample2 reads, "4" is present in example3 reads, "5" exists in exaple4 reads, so after it will be convert to "N" by your principle:

A4:N 
A.: N
A5:N
C4:N
C.: N
C5:N
G4:N
G.: N
G5:N
T4: N
T.:  N
T5: N
N5: A C T or G
N.:  N
N6: N
N0: N
N1: N
N2: N
N3: N

as like that:

@example1
AGGCCAGGATGCATTATGATTACCC
+
611%%(-+%*.&*.,&2,,'%()31
@example2
AGGCCANNNNNNNNNNNNNNNNNNN
+
611%%(-+%*.&*.,&2,,'%()31
@example3
AGGCCAGGATGNNNNNNNNNNNNNN
+
611%%(-+%*.&*.,&2,,'%()31
@example4
AGGCCAGGATGNNNNNGTCGGCAAA
+
611%%(-+%*.&*.,&2,,'%()31

Then you don't need to change the "# SOLiD color code" part in the script. You just need to modify the line 169:

	$current_base = $decode{$colors[$i]}->{$last_base};

change it to:

if (($last_base=~/N/i)&&($colors[$i]==5))
{
	$current_base = $bases[int(rand(@bases))];
}
else
{
	$current_base = (exists $decode{$colors[$i]}->{$last_base}) ? $decode{$colors[$i]}->{$last_base} : "N";
}

It is easier than your ways.

BTW, because SOLiD reads are short, ultra short, most of pepople will abandon these reads which cotain ".456" in color space. I think it is acceptable for SOLiD™ System ultra high throughput, we don't need these uncertain or low quality reads.

[inesdesantiago]

Editing SOLiD2Std.pl to include more colorspace-&gt;basespace

Hi BENM

I edited your SOLid2Std.pl script to include some extra colorspace mapping code that was not considered originally.

I wanted to include the following basespace mapping:
Basically any base(ATCG) that includes a '4' '5' or '.' is 'N'. 'N' to 'N' transition is also represented by diferent color space numbers (0,1,2,3,6,'.').

A4:N 
A.: N
A5:N
C4:N
C.: N
C5:N
G4:N
G.: N
G5:N
T4: N
T.:  N
T5: N
N5: A C T or G
N.:  N
N6: N
N0: N
N1: N
N2: N
N3: N

I am very naive in perl but I tried to change your script to include this conversion.
I edited the following part:

# SOLiD color code
my @code = ([0,1,2,3,'.',4,5],[1,0,3,2,'.',4,5],[2,3,0,1,'.',4,5],[3,2,1,0,'.',4,5],[5,5,5,5,'.',6,0],[5,5,5,5,1,2,3],[5,5,5,5,1,2,3]);
my @bases = qw(A C G T N N N);
my %decode = ();
foreach my $i(0..7)
{
	foreach my $j(0..7)
	{
		$decode{$code[$i]->[$j]} -> {$bases[$i]} = $bases[$j];
	}
}

It works!
However there is an error message when I run the script, although the error does not prevent it from working, which is good.
perl gives me the following error message:

Use of uninitialized value in hash element at SOLid2Std.pl line 49.
Use of uninitialized value within @bases in hash element at SOLid2Std.pl line 49
.

What am I doing wrong?
line 49 is:

$decode{$code[$i]->[$j]} -> {$bases[$i]} = $bases[$j];

Thank You

[nilshomer]

Hi BENM:

Thank you for response with the new information. It happens that I need to convert the SOLiD color space sequence in fastq to Solexa format for its sequence and quality format. I believe the quality score is already in the AscII scheme (see the copied sequence entry in my first email), that is why I thought that that quality score line can be kept without change for my use. Am I right about this? In any case, I think tool for converting among different format of the data from different platform can be useful for us. Thanks again?

Re pliang:

Why would you want to convert color space to sequence space before alignment? Basically, why do you want SOLiD color space data in Illumina format? Bowtie does not work with color space (yet) and no amount of "input hacking" will get it to work right now.

[pliang]

Hi BENM:

Thank you for response with the new information. It happens that I need to convert the SOLiD color space sequence in fastq to Solexa format for its sequence and quality format. I believe the quality score is already in the AscII scheme (see the copied sequence entry in my first email), that is why I thought that that quality score line can be kept without change for my use. Am I right about this? In any case, I think tool for converting among different format of the data from different platform can be useful for us. Thanks again?

[BENM]

Hi BENM,

Thanks for providing the perl script. I am using the SOLiD files from 1000 genome project, and data look like this:

@VAB_Solid0044_20080423_1_Pilot2_YRI_1_8_3KB_MP_11137_718_114
G2203012023131303312303100
+
!611%%(-+%*.&*.,&2,,'%()31

So with your script, the quality line got lost. Just wonder in this case the original quality line can be kept without any change other than removing the first char. I am new to SOLiD data, so want to double check with you. It may be useful for others if you can modify your script to accommodate this format.

Thanks

Hi, pliang

Because samt's question is "Convert SOLiD fastq to Illumina fastq", Illumina FASTQ is different from Standard(Sanger) FASTQ in quality format.

The syntax of Solexa/Illumina read format is almost identical to the FASTQ format, but the qualities are scaled differently. Given a character $sq, the following Perl code gives the Phred quality $Q:

$Q = 10 * log(1 + 10 ** (ord($sq) - 64) / 10.0)) / log(10);

The ASCII charactars in Solexa FASTQ means:

CHAR	DEC	QUALITY
A	65	1
B	66	2
C	67	3
D	68	4
E	69	5
F	70	6
G	71	7
H	72	8
I	73	9
J	74	10
K	75	11
L	76	12
M	77	13
N	78	14
O	79	15
P	80	16
Q	81	17
R	82	18
S	83	19
T	84	20
U	85	21
V	86	22
W	87	23
X	88	24
Y	89	25
Z	90	26
[	91	27
\	92	28
]	93	29
^	94	30
_	95	31
`	96	32
a	97	33
b	98	34
c	99	35
d	100	36
e	101	37
f	102	38
g	103	39
h	104	40
;	59	-5
<	60	-4
=	61	-3
>	62	-2
?	63	-1
@	64	0

In contrast to Solexa FASTQ quality, the ASCII characters in standard (sanger) FASTQ, it used to denote:

CHAR	DEC	QUALITY
!       0       -64
!       1       -63
!       2       -62
!       3       -61
!       4       -60
!       5       -59
!       6       -58
!       7       -57
!       8       -56
!       9       -55
!       10      -54
!       11      -53
!       12      -52
!       13      -51
!       14      -50
!       15      -49
!       16      -48
!       17      -47
!       18      -46
!       19      -45
!       20      -44
!       21      -43
!       22      -42
!       23      -41
!       24      -40
!       25      -39
!       26      -38
!       27      -37
!       28      -36
!       29      -35
!       30      -34
!       31      -33
!       32      -32
!       33      -31
!       34      -30
!       35      -29
!       36      -28
!       37      -27
!       38      -26
!       39      -25
!       40      -24
!       41      -23
!       42      -22
!       43      -21
!       44      -20
!       45      -19
!       46      -18
!       47      -17
!       48      -16
!       49      -15
!       50      -14
!       51      -13
!       52      -12
!       53      -11
!       54      -10
"       55      -9
"       56      -8
"       57      -7
"       58      -6
"       59      -5
"       60      -4
#       61      -3
#       62      -2
$       63      -1
$       64      0
%       65      1
%       66      2
&       67      3
&       68      4
'       69      5
(       70      6
)       71      7
*       72      8
+       73      9
+       74      10
,       75      11
-       76      12
.       77      13
/       78      14
0       79      15
1       80      16
2       81      17
3       82      18
4       83      19
5       84      20
6       85      21
7       86      22
8       87      23
9       88      24
:       89      25
;       90      26
<       91      27
=       92      28
>       93      29
?       94      30
@       95      31
A       96      32
B       97      33
C       98      34
D       99      35
E       100     36
F       101     37
G       102     38
H       103     39
I       104     40
J       105     41
K       106     42
L       107     43
M       108     44
N       109     45
O       110     46
P       111     47
Q       112     48
R       113     49
S       114     50
T       115     51
U       116     52
V       117     53
W       118     54
X       119     55
Y       120     56
Z       121     57
[       122     58
\       123     59
]       124     60
^       125     61
_       126     62
`       127     63
a       128     64

So it is easy to conver Solexa->Sanger quality, you just need to build a conversion table in PERL script, just like this:
# Solexa->Sanger quality conversion table
my @conv_table;
for (-64..64) {
$conv_table[$_+64] = chr(int(33 + 10*log(1+10**($_/10.0))/log(10)+.499));
}

I am trying to write a universal script for Solexa/Illumina, SOLiD/ABi, 454/Roche, 3730/Sanger,...transforming to each other format for different purpose, but I need to know your requirements, after that, I will share it to you all.

Hope I answer your question.
BTW I attach the SOLiD2std.pl for your question, just make a little change in SOLiD2Solexa.pl

[pliang]

Hi BENM,

Thanks for providing the perl script. I am using the SOLiD files from 1000 genome project, and data look like this:

@VAB_Solid0044_20080423_1_Pilot2_YRI_1_8_3KB_MP_11137_718_114
G2203012023131303312303100
+
!611%%(-+%*.&*.,&2,,'%()31

So with your script, the quality line got lost. Just wonder in this case the original quality line can be kept without any change other than removing the first char. I am new to SOLiD data, so want to double check with you. It may be useful for others if you can modify your script to accommodate this format.

Thanks

[BENM]

If your SOLiD FASTQ like this below, you can try this script to convert to Solexa/Illumina FASTQ

@BARB_20071114_2_YorubanMP-BC3_3_16_150_F3
T0220100010131232212020122
+BARB_20071114_2_YorubanMP-BC3_3_16_150_F3
15 21 27 26 24 5 23 18 26 21 11 25 25 19 8 4 25 8 24 7 4 15 18 19 15

[Chipper]

I'm not worried about calling SNPS, just obtaining a consensus sequence mapped to the genome. Would converting to NT from CS still be a bad choice?

Converting will only work as long as you do not have any CS errors (1 CS error will change the rest of the sequence in NT space). The thing with the SOLID is that even with a relatively high CS error rate your mappings will have a very low error rate in NT space due to the 2-base encoding. This is why you need specialised software for the mappings, like BFAST or BWA, else you will get very few alignments.

[nilshomer]

~100 million reads, 34 bps (SOliD), I was hoping to use my machine but I have a powerful enough cluster as well.

It seems you are actively still developing BFAST and there isn't much documentation so its hard to tell if it has all the options I will need for the data. I will want to map, assemble and and align back to the genome.

There is a very complete reference manual that comes with the distribution. Please see download instructions at http://genome.ucla.edu/bfast

[samt]

It's easy to jump off a building, doesn't mean one should.

I'm not worried about calling SNPS, just obtaining a consensus sequence mapped to the genome. Would converting to NT from CS still be a bad choice?

[samt]

Mouse genome is ~3Gb right? In either case, you will have to split the reference to get it to fit on your machine. This is easy and supported with BFAST, although 3Gb is not very much RAM at all. How many reads, what length, and what computational resources do you have?

~100 million reads, 34 bps (SOliD), I was hoping to use my machine but I have a powerful enough cluster as well.

It seems you are actively still developing BFAST and there isn't much documentation so its hard to tell if it has all the options I will need for the data. I will want to map, assemble and and align back to the genome.

[ECO]

It is very easy to do that, maybe I can help you, you can reach me at: [email protected]

It's easy to jump off a building, doesn't mean one should.

[BENM]

I want to do this in order to be able to use Bowtie software, but it doesn't support SOLiD data. Is there software to convert it?

It is very easy to do that, maybe I can help you, you can reach me at: [email protected]