Hi Ateeq,

The reason why you are seeing different sequences for those splice variants
is because you are looking that 5' end which is what differs between the
two splice variants. These two splice variants are transcribed from the
"-" strand. If you take a look at the 3' end you will see that that the
sequences are the same. The other examples you gave are transcribed from
the "+" strand.

I hope that clarifies things for you. If you have further questions, please
email the list: [email protected].

Vanessa Kirkup Swing
UCSC Genome Bioinformatics Group


---------- Forwarded message ----------
From: Ateeq Muhammed Khaliq <[email protected]>
Date: Tue, May 8, 2012 at 3:42 AM
Subject: Re: [Genome] discrepancy in gene co-ordinates
To: "[email protected]" <[email protected]>, "[email protected]"
<[email protected]>, Pauline Fujita <[email protected]>


Hi Pauline,

Thanks a lot for the reply. It's very helpful.

I  have a doubt in the starting sequence range for the two coordinates of
the same gene . Even though we have the same coordinates (Upstream
sequence) in BCL2 gene i.e for chr18:60790579-60986613 and
chr18:60790579-60986657, the UCSC genome browser is showing two different
sequence for the same start regions. Please have a look, It would be great
if you please let me know the Discrepancy or probably, may be i am
interpreting in a wrong way. Kindly clarify.

I have crossed checked the same with NCBI, where it shows common gene
starting sequence for both co-ordinates.( NCBI Reference Sequence:
NG_009361.1 )


Ø  >hg19_knownGene_uc002lit.1 range=chr18:60790579-60986613 5'pad=0 3'pad=0
strand=- repeatMasking=none
TTTCTGTGAAGCAGAAGTCTGGGAATCGATCTGGAAATCCTCCTAATTTT
TACTCCCTCTCCCCGCGACTCCTGATTCATTGGGAAGTTTCAAATCAGCT
ATAACTGGAGAGTGCTGAAGATTGATGGGATCGTTGCCTTATGCATTTGT
TTTGGTTTTACAAAAAGGAAACTTGACAGAGGATCATGCTGTACTTAAAA


Ø  >hg19_knownGene_uc002liu.1 range=chr18:60790579-60986657 5'pad=0 3'pad=0
strand=- repeatMasking=none
CTCGAGCTCTTGAGATCTCCGGTTGGGATTCCTGCGGATTGACATTTCTG
TGAAGCAGAAGTCTGGGAATCGATCTGGAAATCCTCCTAATTTTTACTCC
CTCTCCCCGCGACTCCTGATTCATTGGGAAGTTTCAAATCAGCTATAACT
GGAGAGTGCTGAAGATTGATGGGATCGTTGCCTTATGCATTTGTTTTGGT
TTTACAAAAAGGAAACTTGACAGAGGATCATGCTGTACTTAAAAAATACA


For your reference i am sending you few reference sequences (splice
variants) wherein its perfectly fine.



      I.          VEGFA (uc021yzu.1) at chr6:43737946-43754223 - Homo
sapiens vascular endothelial growth factor A (VEGFA), transcript variant 9,
mRNA.

    II.          VEGFA (uc010jyx.3) at chr6:43737946-43754223 - Homo
sapiens vascular endothelial growth factor A (VEGFA), transcript variant 8,
mRNA.

   III.          VEGFA (uc003owk.3) at chr6:43745460-43754223 - Homo
sapiens vascular endothelial growth factor A (VEGFA), transcript variant 6,
mRNA.

   IV.          VEGFA (uc003owj.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 6, mRNA.

    V.          VEGFA (uc003owi.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 7, mRNA.

   VI.          VEGFA (uc003owh.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 1, mRNA.

 VII.          VEGFA (uc003owg.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 3, mRNA.

VIII.          VEGFA (uc003owf.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 2, mRNA.

   IX.          VEGFA (uc003owe.3) at chr6:43737946-43754223 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 4, mRNA.

     X.          VEGFA (uc003owd.3) at chr6:43737946-43754223 - Homo
sapiens vascular endothelial growth factor A (VEGFA), transcript variant 5,
mRNA.

   XI.          VEGFA (uc003owb.3) at chr6:43737946-43745562 - Homo sapiens
vascular endothelial growth factor A (VEGFA), transcript variant 8, mRNA.

 XII.          VEGFB (uc001nyx.3) at chr11:64002056-64006736 - Homo sapiens
vascular endothelial growth factor B (VEGFB), transcript variant VEGFB-167,
mRNA.

XIII.          VEGFB (uc001nyw.3) at chr11:64002056-64006736 - Homo sapiens
vascular endothelial growth factor B (VEGFB), transcript variant VEGFB-186,
mRNA.

XIV.          PDCD10 (uc003fez.3) at chr3:167401695-167452651 - Homo
sapiens programmed cell death 10 (PDCD10), transcript variant 3, mRNA.

 XV.          PDCD10 (uc003fey.3) at chr3:167401695-167452651 - Homo
sapiens programmed cell death 10 (PDCD10), transcript variant 2, mRNA.

XVI.          PDCD10 (uc003fex.3) at chr3:167401695-167452594 - Homo
sapiens programmed cell death 10 (PDCD10), transcript variant 1, mRNA.


Thanks and Regards,
Ateeq M Khaliq

Bioinformatics Division,
Jubilant Biosys Pvt Ltd,
#96, Industrial Suburb, 2nd Stage
Yeshwantpur, Bangalore 560 022
Ph No : +91-80-66628798


From: Ateeq Muhammed Khaliq
Sent: 08 May 2012 PM 04:12
To: '[email protected]'; '[email protected]'
Subject: RE: discrepancy in gene co-ordinates


Hello all,

This is Ateeq, Sr research Associate, from Jubilant Biosys Ltd, Bangalore
India. I have a doubt regarding amplicon design. While designing an
amplicon for certain gene co-ordinate region, I  have noticed a
discrepancy, which is as follows


http://genome.ucsc.edu/cgi-bin/hgTracks?hgHubConnect.destUrl=..%2Fcgi-bin%2FhgTracks&clade=mammal&org=Human&db=hg19&position=chr18%3A60790579-60986613&hgt.suggest=BCL2&hgt.suggestTrack=knownGene&Submit=submit&hgsid=262656015&knownGene=pack



Ø  For BCL2 gene I got two different nucleotide sequences for the same
start position in UCSC




·        For chr18:60790579-60986657 the  start sequence obtained was-->
CTCGAGCTCTTGAGATCTCCGGTTGGGATTCCTGCGGATTGACATTTCTG

TGAAGCAGAAGTCTGGGAATCGATCTGGAAATCCTCCTAATTTTTACTCC


http://genome.ucsc.edu/cgi-bin/hgc?hgsid=262656015&g=htcDnaNearGene&i=uc002lit.1&c=chr18&l=60790578&r=60986613&o=knownGene&boolshad.hgSeq.promoter=0&hgSeq.promoterSize=1000&hgSeq.utrExon5=on&boolshad.hgSeq.utrExon5=0&hgSeq.cdsExon=on&boolshad.hgSeq.cdsExon=0&hgSeq.utrExon3=on&boolshad.hgSeq.utrExon3=0&hgSeq.intron=on&boolshad.hgSeq.intron=0&boolshad.hgSeq.downstream=0&hgSeq.downstreamSize=1000&hgSeq.granularity=gene&hgSeq.padding5=0&hgSeq.padding3=0&boolshad.hgSeq.splitCDSUTR=0&hgSeq.casing=exon&boolshad.hgSeq.maskRepeats=0&hgSeq.repMasking=lower&submit=submit


And

·        For chr18:60790579-60986613 the start sequence obtained was
TTTCTGTGAAGCAGAAGTCTGGGAATCGATCTGGAAATCCTCCTAATTTT

 TACTCCCTCTCCCCGCGACTCCTGATTCATTGGGAAGTTTCAAATCAGCT

http://genome.ucsc.edu/cgi-bin/hgc?hgsid=262656015&g=htcDnaNearGene&i=uc002liu.1&c=chr18&l=60790578&r=60986657&o=knownGene&boolshad.hgSeq.promoter=0&hgSeq.promoterSize=1000&hgSeq.utrExon5=on&boolshad.hgSeq.utrExon5=0&hgSeq.cdsExon=on&boolshad.hgSeq.cdsExon=0&hgSeq.utrExon3=on&boolshad.hgSeq.utrExon3=0&hgSeq.intron=on&boolshad.hgSeq.intron=0&boolshad.hgSeq.downstream=0&hgSeq.downstreamSize=1000&hgSeq.granularity=gene&hgSeq.padding5=0&hgSeq.padding3=0&boolshad.hgSeq.splitCDSUTR=0&hgSeq.casing=exon&boolshad.hgSeq.maskRepeats=0&hgSeq.repMasking=lower&submit=submit


Can you please explain the reason for the same? Is it an error or I am I
interpreting in other ways? Kindly let me know about this.


Thanks and Regards,
Ateeq M Khaliq

Bioinformatics Division,
Jubilant Biosys Pvt Ltd,
#96, Industrial Suburb, 2nd Stage
Yeshwantpur, Bangalore 560 022
Ph No : +91-80-66628798


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