注册 登录  
 加关注
   显示下一条  |  关闭
温馨提示!由于新浪微博认证机制调整,您的新浪微博帐号绑定已过期,请重新绑定!立即重新绑定新浪微博》  |  关闭

Bioinformatics home

 
 
 

日志

 
 

NGS常用的引物和接头  

2014-03-15 04:59:13|  分类: 默认分类 |  标签: |举报 |字号 订阅

  下载LOFTER 我的照片书  |
http://seqanswers.com/forums/showthread.php?t=198 
Sequences for Solexa Library Preparations: 

Genomic DNA oligonucleotide sequences 

Adapters 1 
5' P-GATCGGAAGAGCTCGTATGCCGTCTTCTGCTTG 
5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT 

PCR Primers 1 
5' AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT 
5' CAAGCAGAAGACGGCATACGAGCTCTTCCGATCT

Genomic DNA Sequencing Primer 
5' ACACTCTTTCCCTACACGACGCTCTTCCGATCT 


DpnII gene expression oligonucleotide sequences 

Gex Adapter 1 
5' P-GATCGTCGGACTGTAGAACTCTGAAC 
5’ ACAGGTTCAGAGTTCTACAGTCCGAC

Gex Adapter 2 
5' CAAGCAGAAGACGGCATACGANN 
5' P-TCGTATGCCGTCTTCTGCTTG 

Gex PCR Primer 1 
5' CAAGCAGAAGACGGCATACGA 

Gex PCR Primer 2 
5' AATGATACGGCGACCACCGACAGGTTCAGAGTTCTACAGTCCGA

Gex Sequencing Primer 
5' CGACAGGTTCAGAGTTCTACAGTCCGACGATC 

NlaIII gene expression oligonucleotide sequences 

Gex Adapter 1 
5' P-TCGGACTGTAGAACTCTGAAC 
5' ACAGGTTCAGAGTTCTACAGTCCGACATG

Gex Adapter 2 
5' CAAGCAGAAGACGGCATACGANN 
5' P-TCGTATGCCGTCTTCTGCTTG 

Gex PCR Primer 1 
5' CAAGCAGAAGACGGCATACGA 

Gex PCR Primer 2 
5' AATGATACGGCGACCACCGACAGGTTCAGAGTTCTACAGTCCGA 

Gex Sequencing Primer 
5' CCGACAGGTTCAGAGTTCTACAGTCCGACATG 


Small RNA oligonucleotide sequences 

RT Primer 
5' CAAGCAGAAGACGGCATACGA 

5' RNA Adapter 
5' GUUCAGAGUUCUACAGUCCGACGAUC 

3' RNA Adapter 
5' P-UCGUAUGCCGUCUUCUGCUUGUidT 

Small RNA PCR Primer 1 
5' CAAGCAGAAGACGGCATACGA 

Small RNA PCR Primer 2 
5' AATGATACGGCGACCACCGACAGGTTCAGAGTTCTACAGTCCGA 

Small RNA Sequencing Primer 
5' CGACAGGTTCAGAGTTCTACAGTCCGACGATC


Genomic DNA

Adapter:
5' -------------------- -----ACACTCTTTCCCTAC ACGACGCTCTTCCGATCT (-) -------------------- -------------- 3'
3' -------------------- -----TGTGAGAAAGGGATG TGCTGCGAGAAGGCTAGp (-) -------------------- -------------- 5'
Adapter:
5' -------------------- -------------------- ------------------ (-) pGATCGGAAGAGCTCGTATG CCGTCTTCTGCTTG 3'
3' -------------------- -------------------- ------------------ (-) TCTAGCCTTCTCGAGCATAC GGCAGAAGACGAAC 5'
PCR Primer:
5' AATGATACGGCGACCACCGA GATCTACACTCTTTCCCTAC ACGACGCTCTTCCGATCT (-) -------------------- -------------- 3' 
3' -------------------- -------------------- ------------------ (-) -------------------- -------------- 5'
PCR Primer:
5' -------------------- -------------------- ------------------ (-) -------------------- -------------- 3'
3' -------------------- -------------------- ------------------ (-) TCTAGCCTTCTCGAGCATAC GGCAGAAGACGAAC 5'
Result Library:
5' AATGATACGGCGACCACCGA GATCTACACTCTTTCCCTAC ACGACGCTCTTCCGATCT (N) AGATCGGAAGAGCTCGTATG CCGTCTTCTGCTTG 3'
3' TTACTATGCCGCTGGTGGCT CTAGATGTGAGAAAGGGATG TGCTGCGAGAAGGCTAGA (N) TCTAGCCTTCTCGAGCATAC GGCAGAAGACGAAC 5'
Genomic DNA Sequencing Primer:
5' -------------------- -----ACACTCTTTCCCTAC ACGACGCTCTTCCGATCT (-) -------------------- -------------- 3'
3' -------------------- -------------------- ------------------ (-) -------------------- -------------- 5'

DpnII gene expression

Gex Adapter 1:
5' -------------------A CAGGTTCAGAGTTCTACAGT CCGAC--------------- -------------------- ------ 3'
3' -------------------- ---CAAGTCTCAAGATGTCA GGCTGCTAGp---------- -------------------- ------ 5'
Gex Adapter 2:
5' -------------------- -------------------- -------------------- ----pTCGTATGCCGTCTTC TGCTTG 3'
3' -------------------- -------------------- -------------------- ---NNAGCATACGGCAGAAG ACGAAC 5'
Gex PCR Primer 1:
5' -------------------- -------------------- ----------------------------------------- ------ 3'
3' -------------------- -------------------- -------------------- -----AGCATACGGCAGAAG ACGAAC 5'
Gex PCR Primer 2:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGA---------------- -------------------- ------ 3'
3' -------------------- -------------------- -------------------- -------------------- ------ 5'
Result Library:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGACGATCNNNNNNNNNNN NNNNNTCGTATGCCGTCTTC TGCTTG 3'
3' TTACTATGCCGCTGGTGGCT GTCCAAGTCTCAAGATGTCA GGCTGCTAGNNNNNNNNNNN NNNNNAGCATACGGCAGAAG ACGAAC 5'
Gex Sequencing Primer:
5' -----------------CGA CAGGTTCAGAGTTCTACAGT CCGACGATC----------- -------------------- ------ 3'
3'--------------------- -------------------- -------------------- -------------------- ------ 5'

NlaIII gene expression

Gex Adapter 1:
5' -------------------A CAGGTTCAGAGTTCTACAGT CCGACATG------------ -------------------- ------ 3'
3' -------------------- ---CAAGTCTCAAGATGTCA GGCTp--------------- -------------------- ------ 5'
Gex Adapter 2:
5' -------------------- -------------------- -------------------- ----pTCGTATGCCGTCTTC TGCTTG 3'
3' -------------------- -------------------- -------------------- ---NNAGCATACGGCAGAAG ACGAAC 5'
Gex PCR Primer 1:
5' -------------------- -------------------- -------------------- -------------------- ------ 3'
3' -------------------- -------------------- -------------------- -----AGCATACGGCAGAAG ACGAAC 5'
Gex PCR Primer 2:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGA---------------- -------------------- ------ 3'
3' -------------------- -------------------- -------------------- -------------------- ------ 5'
Result Library:
5' AATGATACGGCGACCACCGA CAGGTTCAGAGTTCTACAGT CCGACATGNNNNNNNNNNNN NNNNNTCGTATGCCGTCTTC TGCTTG 3'
3' TTACTATGCCGCTGGTGGCT GTCCAAGTCTCAAGATGTCA GGCTGTACNNNNNNNNNNNN NNNNNAGCATACGGCAGAAG ACGAAC 5'
Gex Sequencing Primer:
5' ----------------CCGA CAGGTTCAGAGTTCTACAGT CCGACATG------------ -------------------- ------ 3'
3' -------------------- -------------------- -------------------- -------------------- ------ 5'






Illumina Small RNA sequencing (5' -3'):

Illumina Small RNA Adapter 1                    GTTCAGAGTTCTACAGTCCGACGATC
Illumina Small RNA Adapter 2                    TCGTATGCCGTCTTCTGCTTGT
Illumina Small RNA RT Primer                    CAAGCAGAAGACGGCATACGA
Illumina Small RNA PCR Primer 1                 CAAGCAGAAGACGGCATACGA
Illumina Small RNA PCR Primer 2                 AATGATACGGCGACCACCGACAGGTTCAGAGTTCTACAGTCCGA
Illumina Small RNA Sequencing Primer            CGACAGGTTCAGAGTTCTACAGTCCGACGATC


1.  Adding adaptor
5'  adaptor                                      3'  adaptor  
+ 5'  GUUCAGAGUUCUACAGUCCGACGAUC +++++++++++++ UCGUAUGCCGUCUUCUGCUUGUidT  3'
Reverse transcription                                                           <- 3' AGCATACGGCAGAAGACGAAC 5' 


2. PCR
primer 1: 5'  CAAGCAGAAGACGGCATACGA 3' ->
template  5'  CAAGCAGAAGACGGCATACGA ---------- GATCG TCGGACTGTAGAACTCTGAAC 3'
primer 2:                                                                          <- 3'  AGCCTGACATCTTGAGACTTGGACAGCCACCAGCGGCATAGTAA 5'

3. Small RNA Sequencing                                                                                        
template -      5'    CAAGCAGAAGACGGCATACGA       -------------        GATCG TCGGACTGTAGAACTCTGAACCTGTCGGTGGTCGCCGTATCATT
template +      3'    GTTCGTCTTCTGCCGTATGCT +++++++++++++ CTAGC AGCCTGACATCTTGAGACTTGGACAGCCACCAGCGGCATAGTAA  5'
Sequencing Primer                                                       ++++++++ <-3' CTAGC AGCCTGACATCTTGAGACTTGGACAGC 5' 


 
  评论这张
 
阅读(926)| 评论(0)
推荐 转载

历史上的今天

在LOFTER的更多文章

评论

<#--最新日志,群博日志--> <#--推荐日志--> <#--引用记录--> <#--博主推荐--> <#--随机阅读--> <#--首页推荐--> <#--历史上的今天--> <#--被推荐日志--> <#--上一篇,下一篇--> <#-- 热度 --> <#-- 网易新闻广告 --> <#--右边模块结构--> <#--评论模块结构--> <#--引用模块结构--> <#--博主发起的投票-->
 
 
 
 
 
 
 
 
 
 
 
 
 
 

页脚

网易公司版权所有 ©1997-2017