11 replies to this topic

[zack]

hi admin and chatter,

i need help on dna sequencing analysis

i just got the result

this sequencing are taking at NCBI, code AF012789 for gen cyto B

ORIGIN
1 tttggctctc ttttaggact ctgcttaatt acacaaatcc tcacagggct atttcttgct
61 atacattaca catcagacat ttctaccgcc ttctcatccg tggcacacat ttgccgagac
121 gtaaactatg gatgactaat tcgtaacctc cacgccaacg gcgcctcatt tttctttatt
181 tgtatttatt tccacatcgg ccgaggccta tactacggct cctacctcta taaagagaca
241 tgaaatgtcg gcgtaatact actgctacta gtgatgatga cggcgttcgt agggtacgtt
301 ctaccc
//

this sequence are only 306bp

however last time i test for the same gen cyto B but using this primers

L14841 [5’-AAAAAGCTTCCATCCAACATCTCAGCATGATGAAA-3’] (FORWORD)
and
H15149 [5’-AAACTGCAGCCCCTCAGAATGATATTTGTCCTCA-3’] (REVERSE)

and the result is

> sequence 1 (L14841/FORWORD) are 334bp
AGCTCCTCACTGAGGACTCTGTTTAATCGCACAAATTGTCACAGGGCTATTCCTCGCAATACACTACACGTCTGACATTA
CCACCGCCTTCTCATCCGGAGCCCACATCTGGCGAGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGA
GCCTCTTTCTTCTTTATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATG
AAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCAT
TCTGAGGGGCTGCA

> sequence 1 (H15149/REVERSE) are 347bp
GGCGTTATTTATACATATACCTACAAAGGTGGTTTTATAACTAGTAAGAGAAGAATTACCCCGATGTTTCATGTTTCTTT
ATATAGGTAGGAGCCGTAGTATAGTCCTCGTCCGATGTGGAAATAAATGCAGATAAAGAAAAAAGAGGCTCCGTTGGCGT
GGAGGTTTCGGATTAGTCAACCGTAATTGACGTCTCGGCAGATGTGGGCTACGGATGAGAAGGCGGTGGTAATGTCAGAC
GTGTAGTGTATTGCGAGGAATAGGCCTGTGACAATTTGTGCGATTAAGCAGAGTCCCAGTAGGGAGCCAAAGTTTCATCA
TGCTGAGATGTTGGATGGAAGTTTTAA

> sequence 2 (L14841/FORWORD) are 239bp only
AGGAAAAGAGGAAATTCTGTAGGGGGGCACAAATTGACCAGGCCTATTCCTCGCAATACACTACACGTCTGACATTACCA
CCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCC
TCTTTCTTCTTTATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAA
CATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCATTCT
GAGGGGCTG

> sequence 2 (H15149/REVERSE) are 377bp

ACTCTACATCACAGGAGTTTTTGTAAGAACTATTTTAGAGAAGAATTACCCCGATGTTTCATGTTTCTTTATATAGGTAG
GAGCCGTAGTATAGTCCTCGTCCGATGTGAAAATAAATGCAAATAAAGAAAAAAGAGGCTCCGTTGGCGTGGAGGTTTCG
GATTAGTCAACCGTAATTGACGTCTCGGCAGATGGGGGCTACGGATGAGAAGGCGGTGGTAATGTCAGACGTGTAGTGTA
TTGCGAGGAATAGGCCTGTGACAATTTGTGCGATTAAGCAGAGTCCCAGTAGGGAGCCAAAGTTTCATCATGCTGAGATG
TTGGATGGAAGTTTTAA

> sequence 3 (L14841/FORWORD) are 334 bp

ACAAACTGTATAATCTGTTAGGGGCTCAAATTTACACAGGCCTATTCCTCGCAATACACTACACGTCTGACATTACCACC
GCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTC
TTTCTTCTTTATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACA
TCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCATTCTGA
GGGGCTGCAGTTAA

> sequence 3 (H15149/REVERSE) are 347bp
GNNNNGGGATAACTCCTCTCCTGCTGGGATGGTGTAAGAACTATTTTAGAGAAGAATTACCCCGATGTTTCATGTTTCTT
T
ATATAGGTAGGAGCCGTAGTATAGTCCTCGTCCGATGTGGAAATAAATGCAGATAAAGAAAAAAGAGGCTCCGTTGGCGT
GGAGGTTTCGGATTAGTCAACCGTAATTGACGTCTCGGCAGATGTGGGCTACGGATGAGAAGGCGGTGGTAATGTCAGAC
GTGTAGTGTATTGCGAGGAATAGGCCTGTGACAATTTGTGCGATTAAGCAGAGTCCCAGTAGGGAGCCAAAGTTTCATCA
TGCTGAGATGTTGGATGGAATTTTTA


what's is the problem, all the basepair not the same ... izzit the sequence should be the same as in GENEBANK (NCBI)

[mdfenko]

sequencers make mistakes. you need to get a consensus sequence and compare to that from ncbi.

usually the differences are caused by noise at the beginning and end of the sequence, depending on reaction strength.

keep in mind that sequence data does not usually start at the end of the primer. depending on how you clean the reaction you can start from 5-20 bases after the primer.

Edited by mdfenko, 28 January 2009 - 08:08 AM.

[merlav]

You need a concensus sequence and an aligment tool (ie Omiga, clustal x).

[zack]

here my concensus sequence tree, what the conclusion from that's tree?

[hanming86]

zack, on Jan 29 2009, 11:49 AM, said:

here my concensus sequence tree, what the conclusion from that's tree?

I Know you're from malaysia .

Maybe your DNA template is dirty thus causing subsequent problem either in sequencing or the running of sequencing product.

1. What university are you from?

2. What equipment did you use to check your DNA purity? ( gel can't tell anything about purity , perhaps maybe etoH contamination)

3. And attach your format in .abi format, 1st base is notorious for giving "fake" sequence in the .seq format. A lousy sequence could still generate " good" .seq file if the base calling software is not stringent enough.

4.  what's the first base remark on you seq?

5. Your seq might not be exactly the same as the seq obtained from gen bank unless it's from the exact same source.remember evolution and natural selection.

cheers'

Ming
UTM Skudai

[zack]

hanming86, on Feb 4 2009, 04:18 AM, said:

zack, on Jan 29 2009, 11:49 AM, said:

here my concensus sequence tree, what the conclusion from that's tree?

I Know you're from malaysia .

Maybe your DNA template is dirty thus causing subsequent problem either in sequencing or the running of sequencing product.

1. What university are you from?

2. What equipment did you use to check your DNA purity? ( gel can't tell anything about purity , perhaps maybe etoH contamination)

3. And attach your format in .abi format, 1st base is notorious for giving "fake" sequence in the .seq format. A lousy sequence could still generate " good" .seq file if the base calling software is not stringent enough.

4.  what's the first base remark on you seq?

5. Your seq might not be exactly the same as the seq obtained from gen bank unless it's from the exact same source.remember evolution and natural selection.

cheers'

Ming
UTM Skudai

hi Ming. yes u are right.. i;m from malaysia. and still study at UPM, actually i'm not from molecular background. but i still learning more bout it. the result that;s i send to first base ... i dont know what the conclusion becoz the sample not in the same bp size.

2. i'm using hitachi U2910

3. u can download the attachment

4.

it's my pleasure if u can give some opinion and advice on the .abi format

thank you ming

Attached Files

•  kedah.zip   659.51K   85 downloads

[hanming86]

zack, on Feb 4 2009, 06:16 AM, said:

hanming86, on Feb 4 2009, 04:18 AM, said:

zack, on Jan 29 2009, 11:49 AM, said:

here my concensus sequence tree, what the conclusion from that's tree?

I Know you're from malaysia .

Maybe your DNA template is dirty thus causing subsequent problem either in sequencing or the running of sequencing product.

1. What university are you from?

2. What equipment did you use to check your DNA purity? ( gel can't tell anything about purity , perhaps maybe etoH contamination)

3. And attach your format in .abi format, 1st base is notorious for giving "fake" sequence in the .seq format. A lousy sequence could still generate " good" .seq file if the base calling software is not stringent enough.

4.  what's the first base remark on you seq?

5. Your seq might not be exactly the same as the seq obtained from gen bank unless it's from the exact same source.remember evolution and natural selection.

cheers'

Ming
UTM Skudai

hi Ming. yes u are right.. i;m from malaysia. and still study at UPM, actually i'm not from molecular background. but i still learning more bout it. the result that;s i send to first base ... i dont know what the conclusion becoz the sample not in the same bp size.

2. i'm using hitachi U2910

3. u can download the attachment

4.

it's my pleasure if u can give some opinion and advice on the .abi format

thank you ming

Hey i think your seq can align perfectly well . There seems to be no problem . extra seq flanking the PCR product is expected in sequencing .

I didn't have the time actually to look if the primer seq is within your PCR or not. i need to go eat lunch first now.

And the seq difference from NCBI should be expected if your source is from other individual.

more details on your work would help

k thanks

cheers
-ming

[molgen]

If you run the sequences in ClustalW (or any other alignment tool) you'll see how close the sequences are.
It is very clear that the odd numbers are one type and that the even ones are the other type.

seq4            -----------ACTCTACATCACAGGAGTTTTTGTAAGAACTATTTTAGAGAAGAATTAC 49
seq6            GNNNNGGGATAACTCCTCTCCTGCTGGGATGGTGTAAGAACTATTTTAGAGAAGAATTAC 60
seq2            GGCGTTATTTATACATATACCTACAAAGGTGGTTTTATAACTAGTA-AGAGAAGAATTAC 59
seq3            -------AGGAAAAGAGGAAATTCTGTAGGGGGGCACAAATTGAC-CAG-GCCTATTCCT 51
seq5            ---------ACAAACTGTATAATCTGTTAGGGG-CTCAAATTTACACAG-GCCTATTCCT 49
seq1            -----AGCTCCTCACTGAGGACTCTGTTTAATCGCACAAATTGTCACAG-GGCTATTCCT 54
                                                      ** *     ** *   * *  

seq4            CCCGATGT--TTCATGTTTCTTTATATAGGTAGGAGCCGTAGTATAGTCCTCGTCCGATG 107
seq6            CCCGATGT--TTCATGTTTCTTTATATAGGTAGGAGCCGTAGTATAGTCCTCGTCCGATG 118
seq2            CCCGATGT--TTCATGTTTCTTTATATAGGTAGGAGCCGTAGTATAGTCCTCGTCCGATG 117
seq3            CGCAATACACTACACGTCTGACATTACCACCGCCTTCTCATCCGTAGCCCACATCTGCCG 111
seq5            CGCAATACACTACACGTCTGACATTACCACCGCCTTCTCATCCGTAGCCCACATCTGCCG 109
seq1            CGCAATACACTACACGTCTGACATTACCACCGCCTTCTCATCCGGAGCCCACATCTGGCG 114
                * * **    * ** ** *     **          *        ** ** * ** *  *

seq4            TGAAAATAAATGCAAATAAAGAAAAAAGAGGCTCCGTTGGCGTGGAGGTTTCGGATTAGT 167
seq6            TGGAAATAAATGCAGATAAAGAAAAAAGAGGCTCCGTTGGCGTGGAGGTTTCGGATTAGT 178
seq2            TGGAAATAAATGCAGATAAAGAAAAAAGAGGCTCCGTTGGCGTGGAGGTTTCGGATTAGT 177
seq3            AGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTCTTCTT 171
seq5            AGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTCTTCTT 169
seq1            AGACGTCAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTCTTCTT 174
                 *     ** * *   * *  **     *  ****        ****  *     **  *

seq4            CAACCGTAATTGACGTCTCGGCAGATGGGGGCTACGGATGAGAAGGCGGTGGTAATGTCA 227
seq6            CAACCGTAATTGACGTCTCGGCAGATGTGGGCTACGGATGAGAAGGCGGTGGTAATGTCA 238
seq2            CAACCGTAATTGACGTCTCGGCAGATGTGGGCTACGGATGAGAAGGCGGTGGTAATGTCA 237
seq3            TATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGA 231
seq5            TATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGA 229
seq1            TATCTGCATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGA 234
                 * * * * **     * *  * ** * ** ***       *          **     *

seq4            GACGTGTAGTGTATTGCGAGGAATAGGCCT----GTGACAATTTGTGCGATT--AAGCAG 281
seq6            GACGTGTAGTGTATTGCGAGGAATAGGCCT----GTGACAATTTGTGCGATT--AAGCAG 292
seq2            GACGTGTAGTGTATTGCGAGGAATAGGCCT----GTGACAATTTGTGCGATT--AAGCAG 291
seq3            AACATGAAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTA 291
seq5            AACATGAAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTA 289
seq1            AACATGAAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGTTA 294
                 ** ** *   *   *  *    *   ***    ** * ***    **  **  * *  

seq4            AGTCCCAGTAGGGAGCCAAAGTTTCATCATGCTGAGATGTTGGATGGAAGTTTTAA 337
seq6            AGTCCCAGTAGGGAGCCAAAGTTTCATCATGCTGAGATGTTGGATGGAATTTTTA- 347
seq2            AGTCCCAGTAGGGAGCCAAAGTTTCATCATGCTGAGATGTTGGATGGAAGTTTTAA 347
seq3            TGTATTACCCTGAGGACAAA----TATCATTCTGAGGGGCTG-------------- 329
seq5            TGTATTACCCTGAGGACAAA----TATCATTCTGAGGGGCTGCAGTTAA------- 334
seq1            TGTATTACCCTGAGGACAAA----TATCATTCTGAGGGGCTGCA------------ 334
                 **   *    *  * ****     ***** *****  * **

Edited by molgen, 12 February 2009 - 09:14 AM.

[molgen]

If you Align two sequences (NCBI) an odd number and an even number you'll see that they're the same sequence but frum opiset sides!!!

Length=347

Score =  483 bits (261),  Expect = 4e-141
Identities = 288/300 (96%), Gaps = 5/300 (1%)
Strand=Plus/Minus                                                     <--

Query  2    GCT-CCTCACTGAGGACTCTGTTTAATCGCACAAATTGTCACAGGGCTATTCCTCGCAAT  60
            
Sbjct  307  GCTCCCT-ACTG-GGACTCTGCTTAATCGCACAAATTGTCACAGGCCTATTCCTCGCAAT  250


Query  61   ACACTACACGTCTGACATTACCACCGCCTTCTCATCCGGAGCCCACATCTGGCGAGACGT  120
            
Sbjct  249  ACACTACACGTCTGACATTACCACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGT  190


Query  121  CAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTCTTCTTTATCTG  180
            
Sbjct  189  CAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTTTTCTTTATCTG  130


Query  181  CATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATG  240
            
Sbjct  129  CATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATG  70


Query  241  AAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGT-TATGTAT  299
            
Sbjct  69   AAACATCGGGGTAATTCTTCTCTTACTAGTTATAA-AACCACCTTTGTAGGTATATGTAT  11

Edited by molgen, 12 February 2009 - 09:18 AM.

[zack]

molgen, on Feb 12 2009, 09:11 AM, said:

If you Align two sequences (NCBI) an odd number and an even number you'll see that they're the same sequence but frum opiset sides!!!

Length=347

Score =  483 bits (261),  Expect = 4e-141
Identities = 288/300 (96%), Gaps = 5/300 (1%)
Strand=Plus/Minus                                                     <--

Query  2    GCT-CCTCACTGAGGACTCTGTTTAATCGCACAAATTGTCACAGGGCTATTCCTCGCAAT  60
            
Sbjct  307  GCTCCCT-ACTG-GGACTCTGCTTAATCGCACAAATTGTCACAGGCCTATTCCTCGCAAT  250


Query  61   ACACTACACGTCTGACATTACCACCGCCTTCTCATCCGGAGCCCACATCTGGCGAGACGT  120
            
Sbjct  249  ACACTACACGTCTGACATTACCACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGT  190


Query  121  CAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTCTTCTTTATCTG  180
            
Sbjct  189  CAATTACGGTTGACTAATCCGAAACCTCCACGCCAACGGAGCCTCTTTTTTCTTTATCTG  130


Query  181  CATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATG  240
            
Sbjct  129  CATTTATTTCCACATCGGACGAGGACTATACTACGGCTCCTACCTATATAAAGAAACATG  70


Query  241  AAACATCGGGGTAATTCTTCTCCTACTAGTTATAATAACCGCCTTTGTAGGT-TATGTAT  299
            
Sbjct  69   AAACATCGGGGTAATTCTTCTCTTACTAGTTATAA-AACCACCTTTGTAGGTATATGTAT  11

what that;s mean.. simple explanation pls

Edited by zack, 12 February 2009 - 12:57 PM.

[HomeBrew]

Put all sequences in the same direction by reverse complementing the reverse sequences, and your alignment looks like this:

CLUSTAL 2.0.10 multiple sequence alignment


seq1			---------------------------------------AGCTCCTCACTGAGGACTCTG 21
seq1r		   TTAAAACTTCCATCCAACATCTCAGCATGATGAAACTTTGGCTCCCTACTGGG-ACTCTG 59
seq2			-----------------------------------------AGGAAAAGAGGAAATTCTG 19
seq2r		   TTAAAACTTCCATCCAACATCTCAGCATGATGAAACTTTGGCTCCCTACTGGG-ACTCTG 59
seq3			-------------------------------------------ACAAACTGTATAATCTG 17
seq3r		   -TAAAAATTCCATCCAACATCTCAGCATGATGAAACTTTGGCTCCCTACTGGG-ACTCTG 58
															   *  *   * ****

seq1			TTTAATCGCACAAATTGTCACAGGGCTATTCCTCGCAATACACTACACGTCTGACATTAC 81
seq1r		   CTTAATCGCACAAATTGTCACAGGCCTATTCCTCGCAATACACTACACGTCTGACATTAC 119
seq2			TAGGGGGGCACAAATTGAC-CAGGCCTATTCCTCGCAATACACTACACGTCTGACATTAC 78
seq2r		   CTTAATCGCACAAATTGTCACAGGCCTATTCCTCGCAATACACTACACGTCTGACATTAC 119
seq3			TTAGGGG-CTCAAATTTACACAGGCCTATTCCTCGCAATACACTACACGTCTGACATTAC 76
seq3r		   CTTAATCGCACAAATTGTCACAGGCCTATTCCTCGCAATACACTACACGTCTGACATTAC 118
						* ******  * **** ***********************************

seq1			CACCGCCTTCTCATCCGGAGCCCACATCTGGCGAGACGTCAATTACGGTTGACTAATCCG 141
seq1r		   CACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCG 179
seq2			CACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCG 138
seq2r		   CACCGCCTTCTCATCCGTAGCCCCCATCTGCCGAGACGTCAATTACGGTTGACTAATCCG 179
seq3			CACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCG 136
seq3r		   CACCGCCTTCTCATCCGTAGCCCACATCTGCCGAGACGTCAATTACGGTTGACTAATCCG 178
				***************** ***** ****** *****************************

seq1			AAACCTCCACGCCAACGGAGCCTCTTTCTTCTTTATCTGCATTTATTTCCACATCGGACG 201
seq1r		   AAACCTCCACGCCAACGGAGCCTCTTTTTTCTTTATCTGCATTTATTTCCACATCGGACG 239
seq2			AAACCTCCACGCCAACGGAGCCTCTTTCTTCTTTATCTGCATTTATTTCCACATCGGACG 198
seq2r		   AAACCTCCACGCCAACGGAGCCTCTTTTTTCTTTATTTGCATTTATTTTCACATCGGACG 239
seq3			AAACCTCCACGCCAACGGAGCCTCTTTCTTCTTTATCTGCATTTATTTCCACATCGGACG 196
seq3r		   AAACCTCCACGCCAACGGAGCCTCTTTTTTCTTTATCTGCATTTATTTCCACATCGGACG 238
				*************************** ******** *********** ***********

seq1			AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 261
seq1r		   AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 299
seq2			AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 258
seq2r		   AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 299
seq3			AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 256
seq3r		   AGGACTATACTACGGCTCCTACCTATATAAAGAAACATGAAACATCGGGGTAATTCTTCT 298
				************************************************************

seq1			CCTAC-TAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCAT 320
seq1r		   CTTAC-TAGTTATAAAA-CCACCTTTGTAGGTATATGTATAAATAACGCC---------- 347
seq2			CCTAC-TAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCAT 317
seq2r		   CTAAAATAGTTCTTACA-AAAACTCCTGTGATGTAGAGT--------------------- 337
seq3			CCTAC-TAGTTATAATAACCGCCTTTGTAGGTTATGTATTACCCTGAGGACAAATATCAT 315
seq3r		   CTAAAATAGTTCTTACA-CCATCCCAGCAGGAGAGGAGTTATCCCNNNNC---------- 347
				*  *  ***** * * *	 *	  *							  

seq1			TCTGAGGGGCTGCA----- 334
seq1r		   -------------------
seq2			TCTGAGGGGCTG------- 329
seq2r		   -------------------
seq3			TCTGAGGGGCTGCAGTTAA 334
seq3r		   -------------------

Pretty good agreement -- some minor base calling and end trimming, and you've got your sequence...

[molgen]

Quote

what that;s mean.. simple explanation pls

Zack,
What this means is that you ether did:
T/A clonilg and half went in as 5' to 3' and the other half went in the other way (3' to 5').
Or you sequenced every thing from both sides.

In any case, you have the same sequence every time.

Edited by molgen, 15 February 2009 - 09:16 AM.