tatellano ea UNIVERSAL 100018-USG TELL Seq Library Sequencing

TELL-Seq™ Laeborari ea Tatellano ea Basebelisi
Bakeng sa litsamaiso tsohle tsa tatellano ea Illumina ntle le iSeq 100

Bakeng sa Tšebeliso ea Lipatlisiso Feela. Ha e sebelisoe lits'ebetsong tsa tlhahlobo.
Tokomane # 100018 v8.0
Hlakola 2025

Tokomane ena ke ea mong'a Universal Sequencing Technology Corporation 'me e reretsoe feela ho sebelisoa ke moreki oa eona mabapi le tšebeliso ea lihlahisoa tse hlalositsoeng mona eseng ka mabaka a mang.
Litaelo tse tokomaneng ena li tlameha ho lateloa ka nepo ke basebetsi ba koetlisitsoeng hantle ho netefatsa ts'ebeliso e nepahetseng le e bolokehileng ea lisebelisoa tsa TELL-Seq™.

THEKNOLOJIA YA KA TLHALOSO KA MOKA HA E AKASE MOLAMO MANG KA MANG KA MOR'A TSEBEDISO E fosahetseng ea TELL-SEQ™ KIT.
©2022 Universal Sequencing Technology Corporation. Litokelo tsohle li sirelelitsoe. TELL-Seq™ ke lets'oao la khoebo la Universal Sequencing Technology Corporation. Mabitso a mang kaofela, li-logo le matšoao a mang a khoebo ke thepa ea beng ba tsona.

Selelekela

Protocol ena e hlalosa mokhoa oa ho tsamaisa lilaebrari life kapa life tse nang le indexed-paired TELL-Seq™ tsamaisong ea tatellano ea Illumina®. Laeborari ea TELL-Seq™ † e hloka li-primer tsa tatellano tsa tloaelo bakeng sa sistimi efe kapa efe ea tatellano ea Illumina 'me e na le tatellano ea 18-base index 1 e sebelisoang e le barcode ea molek'hule bakeng sa ho bala ho hokahaneng, e tlamehang ho hlophisoa ka botlalo.

 Litaba tsa Kit

TELL-Seq™ Illumina® Sequencing Primer Kit (PN 100004)

TELL-Seq™ Illumina® Sequencing Primer Kit, HT (PN 100013) 

TELL-Seq™ Sebopeho sa Laeborari le Sekema sa Tatelano

 

Index 1 (ke hore, I7 index) e na le tatelano ea lifaha tsa TELL tse 18, tse LOKELA ho hlophisoa ka botlalo. Index 2 (ke hore, I5 index) e na le 8-base (T-series) kapa 10-base (C-series) sample index primer tatellano e sebelisoang laebraring amplification. Ho khethoa tatellano ea ho qetela e kopanetsoeng. Tlhokahalo e nyane ea bolelele ba ho bala ke 2×96; Bolelele bo boholo bo hlokahalang ke 2×150.

Example ea Illumina® Sequencing % Base by Cycle Chart

Illumina® Sequencing Guideline

1. Fokotsa laeborari ea TELL-Seq™ ho latela maemo le molumo oa sethala sa Illumina®.
2. Lilaebrari li ka kopanngoa hammoho bakeng sa tatellano ha li-primerx tse fapaneng li sebelisoa laeboraring ampmohato oa liification.
3. Li-primaries tsa tatellano tsa tloaelo lia hlokahala ho hlophisa lilaebrari tsa TELL-Seq™ 'me li fanoe ho TELL-Seq™ Illumina Sequencing Primer Kit.
4. Li-primers tsena tsa tatellano ea tloaelo li ka kenngoa ka lilibeng tse boletsoeng bakeng sa li-primers tsa moetlo. Ntle le moo, li ka kenngoa ka lilibeng tse tsamaisanang tsa tatellano ea Illumina® ha laeborari ea taolo ea Illumina® PhiX e ts'oaroa ka tatellano.
5. Custom Index 2 primer e hlokahala feela ha lilaebrari tse ngata tsa TELL-Seq™ tse nang le li-multiplex primers li kopantsoe bakeng sa tatellano le ha sequencer e hloka sekheo sa tatellano ea index ea i5. Hajoale feela bakeng sa MiSeq, tloaelo Index 2 Primer ha e hlokehe. Hape bakeng sa litsamaiso tse sa tšehetsoeng tse kang HiSeq 2000/2500 le NovaSeq v1 reagents, tloaelo Index 2 primer ha e hlokehe.
6. Palo e fokolang ea tatellano ea tatellano e ka etsoa ho sebelisoa palo ea li-primers tsa tatellano tse fanoeng li fapane ho latela sistimi ea tatellano.

Sequencing System	Na ho hlokahala Index 2 Primer?
NovaSeq X/X Plus	Ee
NovaSeq 6000	Ee
HiSeq 3000/4000	Ee
NextSeq	Ee
MiSeq	Che
MiniSeq	Ee

 Example Sample Sheet bakeng sa Sistimi ea MiSeq

Ka tlase ke exampkea tsebaampLe sheet bakeng sa 2 × 146 PE e matha le 18-cycle index 1 (ke hore, I7 index) le 8-cycle kapa 10-base (C-series) index 2 (ke hore, I5 index) ho sebelisa li-primer tsa tatellano ea tloaelo bakeng sa Bala 1 , Index 1 le Read 2 e tla kenngoa ka har'a liliba tsa tatellano ea tloaelo. Hobane MiSeq e sebelisa primer ea P5 holim'a selefouno ea phallo ha tatellano ea tatellano ea Index 2 e bala, TELL-Seq™ custom Index 2 primer ha e hlokehe bakeng sa tsamaiso ea MiSeq. Ho fokotseha ha lilaebrari life kapa life tse kopantsoeng ho tla theoa holim'a sample index (ke hore, index 2). E tla sebetsoa ka mokhoa oa TELL-bala oa tlhahlobo ka thoko ka mor'a hore tatellano e phethe.

 

Illumina® Sequencing Bala Bolelele Keletso

1. Ho kgothaletswa hore ho be le tatellano ya bofelo e kopanetsweng.
2. TELL-Seq™ library Index 1 ke 18-base, Index 2 ke 8-base (T-series) kapa 10-base (C-series). Ho na le kakaretso ea 26-base (T-series) kapa 28-base (C-series) bakeng sa li-index ka bobeli ha li bapisoa le kakaretso ea 16-base bakeng sa index e tloaelehileng ea Illumina dual.
3. Li-cycle tse ling tse 10 tse hlokahalang bakeng sa tatellano ea index ea laeborari ea TELL-Seq™ e hloka ho tlosoa ho bala 1 le ho bala lipotoloho tse 2 ka ho lekana. Kaha Illumina sequencing reagent e tiisa lipotoloho tse 2 tse eketsehileng, 4-cycle bakeng sa ho bala 1 le 4-cycle bakeng sa ho bala 2 e hloka ho tlosoa ka ho latellana.
4. Bolelele ba tatellano bo khothalelitsoeng ke 2 × 96 PE ho 2 × 146 PE bakeng sa li-index tse peli; 2×100 PE ho isa ho 2×150 PE bakeng sa s e le 'ngoeample matha ntle le tlhokahalo ea ho bala Index 2.

Tatellano Tebileng ho Nahana

1. Ho hlokahala botebo bo lekaneng ba tatellano ho fumana tšireletso e lekaneng ea TELL Bead. Ha TELL Bead e sebelisoa laeboraring amplification ho hlahisa laeborari ea TELL-Seq™, ho tla hlokahala hore ho baloe ka tatellano e eketsehileng ho fumana botebo bo batloang ba tatellano. Leha ho le joalo, TELL Bead tse fokolang tse sebelisoang laeboraring amplintlheng, ho tla ba tlase ho rarahana ha laebrari, e leng se ka lebisang ho sekhahla se phahameng sa ho pheta-pheta ha ho bala ka tatellano. Ho leka-lekana lipakeng tsa TELL Beads tse sebelisitsoeng le ho rarahana ha laeborari ea TELL-Seq™ ho ka ipapisa le boholo ba genome le ts'ebeliso.
2. Bakeng sa ts'ebeliso ea kopano ea de novo, bonyane 60x ts'ebetso ea genome ea sample kgothaletswa ka kakaretso. Leha ho le joalo, khaso e tlase ea tatelano e kanna ea lekana ho latela palo ea TELL Beads e sebelisoang laeboraring amplification le ho rarahana ha laeborari ea TELL-Seq™.
3. Bakeng sa phaliso ea batho, bonyane lihlopha tse limilione tse 500 li baloa ka k'hothomoampLe e leng ~ 40x botebo bo khothalelitsoeng.
4. Bakeng sa tatellano e reriloeng, ho khothaletsoa ho fana ka bonyane 100x.

Laebrari Multiplex Primer Index Sequences (ke hore, Index 2 Bala): T-series (8-base)

Library Multiplex Primer (T-letoto)	Bakeng sa Sample Sheet MiSeq	Bakeng sa Sample Sheet NovaSeq, NovaSeq X, NextSeq, MiniSeq
T501	TGAACCTT	AAGGTTCA
T502	TGCTAAGT	KETSAHALOTAGCA
T503	TGTCTCT	AGAGACA
T504	TAAGACAC	GTGTCTTA
T505	CTAATCGA	TCGATTAG
T506	CTAGAACA	TGTTCTAG
T507	TAAGTTCC	GGAACTTA
T508	TAGACCTA	TAGGTCTA
T509	CATCCGAA	TTCGGATG
T510	TTATGAGT	ACTCATAA
T511	AGAGGCGC	GCGCCTCT
T512	TAGCCGCG	CGCGGCTA
T513	ACGAATAA	TTTTTGT
T514	TTCGTAGG	CTCTACGAA
T515	GATTGCT	AGCAGATC
T516	CGCTCCGC	GCGGAGCG
T517	AGGCTATA	TATAGCCT
T518	GCCTCTAT	ATAGAGGC
T519	AGGATAGG	KHOTHATSO
T520	TCAGAGCC	GGCTCTGA
T521	CTTCGCCT	AGGCGAAG
T522	TAAGATTA	TAATCTTA
T523	AGTAAGTA	TS'OANELO
T524	GACTTCCT	AGGAAGTC

 Laebrari Multiplex Primer Index Tatelano (ke hore Index 2 Bala): C,D,E,F-series (10-base)

Library Multiplex Primer (C-letoto)	Bakeng sa Sample Sheet MiSeq	Bakeng sa Sample Sheet NovaSeq, NovaSeq X, NextSeq, MiniSeq
C501	ACGTACGTAC	GTACGTACGT
C502	CATGCATGCA	TGCATGCATG
C503	GTACGTACGT	ACGTACGTAC
C504	TGCATGCATG	CATGCATGCA
C505	ATGCTGATCA	TGATCCAGAT
C506	CACAGCTGTG	CACAGCTGTG
C507	GCTGATCAGC	GCTGATCAGC
C508	TGATCCAGAT	ATGCTGATCA
C509	TS'OATSOE	AGTATTGAAT
C510	CTAGCGCTAG	CTAGCGCTAG
C511	GCTAGTAGTA	TACTACTAGC
C512	TCCAATCAAG	CTTGATTGGA
C513	AATATTGCTG	CAGCAATATT
C514	CGTCGTTACG	CGTAACGACG
C515	GATTGATTCC	GGAATCAATC
C516	TCTAACAATG	CATTGTTAGA
C517	AGAATTGTCA	TGACAATTCT
C518	CTCAGCAATT	AATGCTGAG
C519	GGTCCTTGTC	GACAAGGACC
C520	AGGCCTGACA	TGTCAGGCCT
C521	CTCCTAGTGG	CCACTAGGAG
C522	GGTTACAGCT	AGCTGTAACC
C523	CTGATTGGCG	CGCCAATCAG

C524	ATTGGTTAGA	TS'ELISITSOE
C525	KHOTHATSO	AGTTGAATGG
C526	CAGTATTGAC	GTCAATACTG
C527	GAGTCCTCAA	TTGAGGACTC
C528	AGCTACTACT	AGTAGTAGCT
C529	TAGCTAGCGC	GCGCTAGCTA
C530	GATGCACAC	GTGTTGCATC
C531	CCTCAGTACA	TGTACTGAGG
C532	CGGATATTCA	TGAATTACCG
C533	CGCAATGGCT	AGCCATTGCG
C534	GTACGTTGAA	TTCAACGTAC
C535	TTGATCAGTA	TACTGATCAA
C536	GGCCTAACAA	TTGTTAGGCC
C537	GTTGTTGGAA	TTCAACAAC
C538	TACGTTGGAC	GTCCAACGTA
C539	ACACCATGCA	TGCATGGTGT
C540	GCAATAGTAC	GATATTGC
C541	ACGCAGCCAG	CTGGCTGCGT
C542	CGAGTTGACG	CGTCACTCG
C543	CGTGGCTGAA	TTCAGCCACG
C544	TCTCAAGGAC	GTCCTTGAGA
C545	CCTAGGCACT	AGTGCCTAGG
C546	CTGCGGTAAT	ATTACCGCAG
C547	GGCACTACCA	TGGTAGTGCC
C548	GCTCAATCAA	TTGATTGAGC
C549	AGGCACACAC	GTGTGTGCCT
C550	CCTGGCAAGA	TCTTGCCAGG

C551	TATTGGTAG	CATACAATTA
C552	GCCAACAAGT	ACTTGTTGGC
C553	ATGGCTTATA	TATAAGCCAT
C554	GCATGGCCTT	AAGGCCATGC
C555	ACAATACTGG	CCAGTATTGT
C556	GGATTGGACT	AGTCCAATCC
C557	ACTGTACTAT	ATAGTACAGT
C558	CAGCTGTGAG	CTCAAGCTG
C559	CTTGAGGACC	GGTCTCCAAG
C560	GGTACAATAG	CTATGTACC
C561	CTGACTACTA	TAGTAGTCAG
C562	TCAACCATGG	CCATGGTTGA
C563	ATATAACCG	CGGTTATAAT
C564	ACTAGTCCTT	AAGGACTAGT
C565	ACTTGGACGT	ACGTCCAAGT
C566	ATGGTTAGGA	TCCAACCAT
C567	ATGGTACCAA	TTGGTACCAT
C568	GAATTGACTC	GAGTCAATTC
C569	AGCAACCAGG	CCTGGTTGCT
C570	TACTGTGCTG	CAGCACAGTA
C571	CAACAACGTC	GACGTTGTTG
C572	CAGTAGCGCT	AGCGCTACTG
C573	ATTACCAATC	GATTGGTAAT
C574	TAAGGACCGC	GCGTCCTTA
C575	ACACGTACCG	CGGTACGTGT
C576	CAAGTTGTT	AACAACGTTG
C577	ATTGTGCTGA	TEKACACAAT

C578	GTACAACAG	CTGTTGGTAC
C579	TTGTCAAGGA	TCCTTGACAA
C580	CTTGTACGTA	TACGTACAAG
C581	TGCCTTGTAA	TTACAAGGCA
C582	TAGTAGCTTA	TAAGCTACTA
C583	GACCCAATG	CATTGCGGTC
C584	CTACTAGCTT	AAGCTAGTAG
C585	AGCACGTT	AACGTGTGCT
C586	TGTTATAAGC	GCTTATAACA
C587	GTTGCCAAGT	ACTTGGCAAC
C588	CTGCAACCG	CGGTTGCCAG
C589	TTAGGCCTTA	TAAGGCCTAA
C590	CGCAGCACAG	CTGTGCTGCG
C591	CTAGGCCAA	TTGTGCCTAG
C592	TGTTGTACAG	CTGCAAACA
C593	CTACGTGGC	GCACGTTAG
C594	GCGTACTGGT	ACCAGTACGC
C595	GGCCTGAATT	AATTCAGGCC
C596	CATGCTCGAG	CTCGAGCATG

Library Multiplex Primer (D-letoto)	Bakeng sa Sample Sheet MiSeq	Bakeng sa Sample Sheet NovaSeq, NovaSeq X, NextSeq, MiniSeq
D501	AGCACTGTAA	TTACAGTGCT
D502	CCGAAGTACT	AGTACTTCGG
D503	GTTCAAGTA	TACTTGTAAC
D504	TGATTGCGTG	CACGCAATCA
D505	ATCTTAGGCA	TGCCTAAGAT

D506	CACGCAAGTT	AACTTGCGTG
D507	GAGCTTAGGA	TCCTAAGCTC
D508	TATTGCTCGA	TCGAGCAATA
D509	AACATCTGAG	CTCAGATGTT
D510	CAACTCAGCA	TGCTGAGTTG
D511	GAACTATATG	CATATAGTTC
D512	TAATAGTCTA	TAGACTATTA
D513	AAGATTGACG	CGTCAATCTT
D514	CACGTAGCCG	CGGCTACGTG
D515	GACTCGATCA	TGATCGAGTC
D516	TAATCCTCGC	GCGAGGATTA
D517	AAGCTCCTGG	CCAGGAGCTT
D518	CAGATTGTTG	CACACAACTG
D519	GATGCTATTC	GAATAGCATC
D520	TAATCCTCGC	GCGAGGATTA
D521	AAGATACCT	AGGTATACTT
D522	CAGTAGCCAA	TTGGCTACTG
D523	GCAAGGACTC	GAGTCCTTGC
D524	TACTGTA	TACAGTAGTA
D525	AATCCTAGCT	AGCTAGGATT
D526	CATTGGCAAG	CTTGCCAATG
D527	GCCTGAGAAT	ATTTCAGGC
D528	TACTAGCAAG	CTTGCTAGTA
D529	ACAGCTAGTC	GACTAGCTGT
D530	CCAACCAGTA	TACTGGTTGG
D531	GCTCAATT	AATTGAGCGC
D532	TAGCGCGGAC	GTCCGCGCTA

D533	ACCTATAGTT	AACTATAGGT
D534	CCCGTCAGG	CCTGACGTGG
D535	GCTACGCAAT	ATTGCGTAGC
D536	TAGTTGCTGC	GCAGCAACTA
D537	ACGGTACAAC	GTTGTACCGT
D538	CCGAACTTCG	CGAAGTTCGG
D539	GCTCCTAATT	AATTAGGAGC
D540	TATCCTCGAT	ATCGAGGATA
D541	ACTCGATCT	AGAATCGAGT
D542	CCTGGTAATT	AATTACCAGG
D543	TCGTAGTTGG	CCAACTAAGC
D544	TCAGCAAGGA	TCCTTGCTGA
D545	ACTGGTAGCA	TGCTACCAGT
D546	CGATCACCT	AGGTGATCG
D547	GGAAGTAGCA	TGCTACTTCC
D548	TCAGCTGCGG	CCGCAGCTGA
D549	ACTGGTAGCA	TGCTACCAGT
D550	CGATCACCT	AGGTGATCG
D551	GGCAAGATGA	TCATCTTGCC
D552	TCCGCCAATG	CATTGGCGGA
D553	AGAACTTAT	ATAAGATTCT
D554	CGCTAGCAAC	GTTGCTAGCG
D555	GGCCTTATAT	ATATAAGGCC
D556	TCGAATACTA	TAGTATTCGA
D557	AGCAATGGTA	TACCATGCT
D558	CGGTCTGATC	GATCAGACCG
D559	GGTCGCCCAT	ATGGCGGACC

D560	TCGAATACTA	TAGTATTCGA
D561	AGCACTGTAA	TTACAGTGCT
D562	CGTACTACCG	CGGTAGTACG
D563	GGTTCATCGG	CCGATGAACC
D564	TCGGAGCTGC	GCAGCTCCGA
D565	AGGACGTTAG	CTAACGTCCT
D566	CGTTGACAAT	ATTGTCAACG
D567	GTACTGACAC	GTGTCAGTAC
D568	TCTAAGATAG	TATATAGA
D569	AGTGCTCAAC	GTTGAGCACT
D570	CTAAGAACCT	AGGTCTTAG
D571	GATAATTAC	GTAATATAC
D572	TGATTGCGTG	CACGCAATCA
D573	ATTCGACG	CGTCGAATAT
D574	CTATAACGAC	GTCGTTATAG
D575	GTCGTTAGGT	ACCTAACGAC
D576	TGCCACTGAT	ATCAGTGGCA
D577	ATCGGCTAAG	CTTAGCCGAT
D578	CTCAAGTTGC	GCAACTTGAG
D579	GTGTGTTCGA	TCGAACACAC
D580	TGCTATAGTC	GACTATAGCA
D581	ATGAGCTATT	AATAGCTCAT
D582	KHOTHATSO	TCGTTGTGAG
D583	TGCTCGATTG	CAATCGAGCA
D584	ATGTTGGACT	AGTCCAACAT
D585	CTGATGTTAG	CTACATCAG
D586	TGGTAACAG	CTGGTTACCA

D587	ATTCGAAGTA	TACTTCGAAT
D588	CTTAGCTCCT	AGGAGCTAAG
D589	TGTAATCGAT	ATCGATTACA
D590	CTTGGACTTC	GAAGTCCAAG
D591	TGTACTTGAA	TTCAAGTACA
D592	TTAAGCTCAG	CTGAGCTTAA
D593	TTACAACCAG	CTGTTGTAA
D594	TCTCAGCCA	TGGCTGAGAA
D595	TTGCATGGCT	AGCCATGCAA
D596	TTGGCGGTAC	GTACCCGCCAA

Library Multiplex Primer (E-letoto)	Bakeng sa Sample Sheet MiSeq	Bakeng sa Sample Sheet NovaSeq, NovaSeq X, NextSeq, MiniSeq
E501	AGCCCGGTC	GACCGTGGCT
E502	CCGCCTGAGT	ACTCAGGCGG
E503	GTTATATACC	GGATATAAC
E504	TTCCTGGCA	TGCCAGGAAG
E505	ATCGAATTGC	GCAATTCGAT
E506	CACGTGTTGT	ACAACACGTG
E507	GAGCTGTGTT	AACCAGCTC
E508	TATACAGAT	ATCTGTAATA
E509	ACAATGGCG	CGCATTGTT
E510	CAACTCGCTA	TAGCGAGTTG
E511	GAATTGCCTCA	TGAGCAATTC
E512	GTGTGTTCGA	TCGAACACAC
E513	CGATTCGAGC	GCTCGAATCG
E514	GTCAACGTTA	TAAGTTGAC

E515	GACAATCCTA	TAGGATTGTC
E516	TATCAGACT	AGTCTGATTA
E517	AAGCTATAT	ATATAAGCTT
E518	CATATTGTAA	TTACAATATG
E519	GATAGGACTT	AAGTCCTATC
E520	ATGCTAGGTT	AACCTAGKATSE
E521	GCAAGATTAG	CTAATCTTGC
E522	CAGCTGTATA	TATACAGCTG
E523	GCAAGTCTAA	TTAGACTTGC
E524	TACTAGCAAG	CTTGCTAGTA
E525	AATACTGTTA	TAACAGTATT
E526	CATATCTGAC	GTCAGATATG
E527	GCACTAAGG	CCTTAGTGGC
E528	CTGGCAATTC	GATTGCCAG
E529	ACAATAGGCG	CGCCTATTGT
E530	CCATGCTAAG	CTTAGCATGG
E531	GCGAGCGATC	GATCGCTCGC
E532	TAGGTAGTTC	GAACTACTA
E533	ACCTTGGACC	GGTCCAAGGT
E534	TGACTAATAC	GTATTAGTCA
E535	GCTATATCTG	CAGATATAGC
E536	TCTAGTTCAG	CTGAACTAGA
E537	ACGGTTGAGA	TCTCAACCGT
E538	CTGATTCCGG	CCGGAATCAG
E539	TGGTAACAG	CTGGTTACCA
E540	GACTCAGACA	TGTCTGAGTC
E541	TTAGCCAATA	TATTGGCTAA

E542	CCTAAGGCG	CGCCTTAAGG
E543	CGTAGCAATC	GATTGCTACG
E544	KHABANE	ATTAGCCATG
E545	ACTCGACAAT	ATTGTCGAGT
E546	CGAGTCTAGG	CCTAGACTCG
E547	GGACTCAGCT	AGCTGAGTCC
E548	TCAGCAAGGA	TCCTTGCTGA
E549	CGATTCTTA	TAAGGAATCG
E550	ATTAGGCAAT	ATTGCCTAAT
E551	TTGTCGAAGG	CCTTCGACAA
E552	TCCAAGTAGC	GCTACTTGGA
E553	AGATAGTCCG	CGGACTATCT
E554	CGCTAACATA	TATGTTAGCG
E555	GGCTAATTGT	ACAATTAGCC
E556	AGGTCGTCA	TGACGTACCT
E557	TTCCAAGTTC	GAACTTGGAA
E558	CGGAGCTCCG	CGGAGCTCCG
E559	GGTACTAATA	TATATAGTACC
E560	TCGAAGGCAA	TTGCCTTCGA
E561	CTCCTTGATG	CATCAAGGAG
E562	GCAATTCCGG	CCGGAATTGC
E563	ACTACCAGC	GCTGCGTAGT
E564	AGGAATGGCC	GGCCATTCCT
E565	AGGTTAGKATSE	ATGCTAACCT
E566	CGTCCTAGAC	GTCTAGGACG
E567	GCGTGCAAT	ATTGCCACGC
E568	TCTGATCGAC	GTCGATCAGA

E569	AGTTGTCAGG	CCTGACAACT
E570	CTACGCCAAT	ATTGCGTAG
E571	GTAGTTGCGT	ACGCAACTAC
E572	TGAGCGCTAT	ATAGCGCTCA
E573	ATACCTTAGT	ACTAAGGTAT
E574	GCACCATT	AATGGTGTGC
E575	GTCTAAGGAG	CTCCTTAGAC
E576	CATTCCCGGG	CCGCGGAATG
E577	GCGTACCTAG	CTAGGTACGC
E578	CTCAATAGCA	TGCTATTGAG
E579	GTGCTAAGCC	GGCTTAGCAC
E580	TGCCACTATC	GATAGTGGCA
E581	GCTGTTGAAT	ATTCAACAGC
E582	CGGCTAATCC	GGATTAGCCG
E583	AACTACCGCA	TGCGGTAGTT
E584	ATGCATCAAG	CTTGATGCAT
E585	CTGACTTGCA	TGCAAGTCAG
E586	TGGTACCCTAT	ATAGGTACA
E587	ATTAGCTAAC	GTTAGCTAAT
E588	TGGTACCCTAT	ATAGGTACA
E589	TGTTGCTACC	GGTAGCAACA
E590	CTTATATAAG	CTTATATAAG
E591	TCAGTCTGT	ACAGACTTGA
E592	TGAGCGCTAT	ATAGCGCTCA
E593	TTAGGCGTAC	GTACGCCTAA
E594	TCTCATCAG	CTGATGAGAA
E595	CTAGGTATTG	CAATACCTAG

E596

TTGCTGACTA

TAGTCAGCAA

Library Multiplex Primer (F-letoto)	Bakeng sa Sample Sheet MiSeq	Bakeng sa Sample Sheet NovaSeq, NovaSeq X, Next Seq, MiniSeq
F501	AGCCAGTAGC	GCTACTGGCT
F502	CCGATGAGTT	AACTCATCGG
F503	GTTCAACTTC	GAAGTTGAAC
F504	TTGAGCATCA	TGATGCTCAA
F505	ATCACAGTG	CACTGTTGAT
F506	CACACGGTAG	CTACCGTGTG
F507	GAGGATTGCT	AGCAATCCTC
F508	TATCTTGGCC	GGCCAAGATA
F509	AACTAGTGGC	GCACTAGTT
F510	CAATCTGGT	ACCAGGATTG
F511	GAAGGTTACG	CGTAACCTTC
F512	CTAAGAACCT	AGGTCTTAG
F513	CACACGGTAG	CTACCGTGTG
F514	AATTAGTCTG	CAGACTAATT
F515	GACCGTGTTA	TAACCGGTC
F516	TATTGAGG	CCTCGAATTA
F517	AAGATTGACG	CGTCAATCTT
F518	CAACTCGCTA	TAGCGAGTTG
F519	GATTCGAACT	AGTTCGAATC
F520	GCCTTGAGTT	AACTCAAGGC
F521	CACAGATTAG	CTATCTGTG
F522	CAGCTAGGCC	GGCCTAGCTG
F523	GCAAGGTATA	TATACCTTGC

F524	TACCTAGGTA	TACCTAGGTA
F525	AATCCTTGAA	TTCAAGGATT
F526	KHABANE	AGGTTCGATG
F527	GCCTGCGTAA	TTACGCAGGC
F528	KHABANE	AGGTTCGATG
F529	ACAGTGCCAT	ATGGCACTGT
F530	CCAACTATTG	CAATAGTTGG
F531	GCGTACCGCA	TGCGGTACGC
F532	TAGGAGCGAT	ATCGCTCCTA
F533	ACCAATTGGT	ACCAATTGGT
F534	AATACTGTTA	TAACAGTATT
F535	GCTGTTGAAT	ATTCAACAGC
F536	TTACAACCAG	CTGTTGTAA
F537	ACGTCAAGTC	GACTTGACGT
F538	KHOTHATSO	CATTGAATTG
F539	TGAATTCAGC	GCTGAATTCA
F540	ACGTCAGCT	AGCTGTACGT
F541	TGACTACCAT	ATGGTAGTCA
F542	CCTGGTGTGA	TCACACCAGG
F543	GACTAATCGC	GCGATTAGTC
F544	ACGTTAGTGC	GCACTAACGT
F545	ACTCGATCT	AGAATCGAGT
F546	CGACAAGATT	AATCTTGTCG
F547	GGAATTGGCT	AGCCAATTCC
F548	TCAAGCCGA	TCGGCTGTGA
F549	CTGTACTGTA	TACAGTACAG
F550	TCGCTCGAGG	CCTCGAGCGA

F551	ACCTTAGCAA	TTGCTAAGGT
F552	TCCGGACTAC	GTAGTCCGGA
F553	AGAACTTGCA	TGCAAGTTCT
F554	CGCAGGACTT	AAGTCCTGCG
F555	GGCAAGATGA	TCATCTTGCC
F556	GCCTAGCGCG	CGCGCTAGGC
F557	GACATCGCTA	TAGCGATGTC
F558	CGGCTAATCC	GGATTAGCCG
F559	GGTTGAGCTT	AAGCTCAACC
F560	TCGCTCAGGC	GCCTGAGCGA
F561	GCTGATTATA	TATAATCAGC
F562	GCGCTAGTCC	GCACTAGCGC
F563	TATGTCTGAA	TTCAGACATA
F564	AAGCTATAT	ATATAAGCTT
F565	AGGAATGGCC	GGCCATTCCT
F566	CGTAGCGCGC	GGCGCTACG
F567	GCCTGCGTAA	TTACGCAGGC
F568	TCTTAGCGAT	ATCGCTAAGA
F569	AGTTCTAAGA	TCTTAGAACT
F570	CTAGGTATTG	CAATACCTAG
F571	GTAGTACTAC	GTAGTACTAC
F572	TGACAAGTAG	CTACTTGTCA
F573	ATAGTCTGAG	CTCAGACTAT
F574	CTGTGTTCA	TGTAACACAG
F575	GTCTAGCCAC	GTGGCTAGAC
F576	AATCAAGGTT	AACCTTGATT
F577	CATAATTATG	CATAATTATG

F578	CTCGAGCTAT	ATAGCTCGAG
F579	GTGCCTGATG	CATCAGGCAC
F580	TGCTCGATTG	CAATCGAGCA
F581	GATTCGAACT	AGTTCGAATC
F582	TCGTTGAGGA	TCTCAACGA
F583	AGCCAGGCGT	ACGCCTGGCT
F584	ATGTCAGATT	AATCTGACAT
F585	CTGATATAC	GATATACAG
F586	TGGTGTCAAC	GTTGACACCA
F587	ATTGCATGCT	AGCATGCAAT
F588	AGTATTCATA	TATGAATACT
F589	TGTAATTCCG	CGGAATTACA
F590	TTCCTGGCA	TGCCAGGAAG
F591	TCCGCGGA	TCCGCGGA
F592	CACTGTCGG	CCGAACAGG
F593	TTAGCCAATA	TATTGGCTAA
F594	TTCGAGCTGA	TCAGCTCGAA
F595	GTACCGCGCG	CGCGCGGTAC
F596	TTGAGCATCA	TGATGCTCAA

Sehlomathiso

Li-Priming TELL-Seq™ Custom Primers ho li-Prime tsa Tatelano tsa Illumina®

Lilaebrari tsa TELL-Seq™ li hloka li-primer tsa tatellano tse ikhethileng bakeng sa li-platform tsa tatellano ea Illumina. Spiking (kapa ho kopanya) TELL-Seq™ Illumina tatellano ea tatellano ea li-primers ho li-primer tsa tatellano tsa Illumina hoa hlokahala ha ho kenyelletsoa laeborari ea taolo ea PhiX kapa lilaebrari tse ling tse tloaelehileng tsa Illumina tse nang le lilaebrari tsa TELL-Seq™ ka tatellano. Tlhokomeliso: TELL-Seq™ Index 1 e bala e na le 18-base mme e hloka tatellano ea li-cycle 18; Index 2 bala e na le 8-base bakeng sa T-series primer le 10-base bakeng sa C-series primer. Mokhoa oa ho kenya li-primer tsa tloaelo ho li-primer tsa Illumina

• Boemo ba cartridge, palo e felletseng, le khatello ea ho qetela ea li-primers tsa sethala ka seng li fanoe ka har'a litafole tse ka tlase.
• Bala bophahamo ba molumo oa primer e tloahelehileng ho e eketsa sebakeng sa Illumina primer cartridge ho ipapisitse le boemo ba ho qetela ba premier ea tloaelo ka har'a khatriji.
• Ka mor'a hore u phunyeletse setsing sa tloaelo, fetola pipette ho halofo ea molumo o feletseng 'me ka bonolo pipette holimo le tlaase ka makhetlo a mahlano ho kopanya ka botlalo.

Bakeng sa li-platform tsa MiSeq, MiniSeq, NextSeq le NovaSeq, u se ke oa hlahloba boemo ba lebokose la "custom primer" ho s.ampletlapa kapa nakong ea ho seta. * Ho bala hore na ho na le tokomane e kae e ka kenang selibeng, sebelisa (C1)*(V1) = (C2)*(V2) equation moo:

• V1 = rarolla hore molumo oa primer ea tloaelo e kenngoe ho eona
• C2 = tlhahiso ea ho qetela e khothaletsoang ho tloha chate e ka tlase
• V2 = kakaretso ea boholo ba Illumina primer lichabeng tse ka tlase

Example bakeng sa sethala sa MiSeq:
100M * V1 = 0.5M * 680L V1 = 3.4 µl

Keletso ea Bohlokoa: Litataiso tse ka tlase li ipapisitse le meqolo ea hajoale ea li-primer. Ho etsa bonnete ba ho nepahala, lekanya palo eohle ea li-primer ka har'a cartridge ka pipette pele u tsoela pele ka ho seta.

Sequencer e 'ngoe le e' ngoe ea Illumina® e na le mahloriso a fapaneng a ho qetela le molumo o felletseng kahoo sheba tokomane e ka tlase ho netefatsa hore palo e nepahetseng ea TELL-Seq™ Illumina® Sequencing Primers e eketsoa ka kotloloho.

*Tlhokomeliso: Index 1 le Index 2 hammoho le Bala 1 le Bala 2 Li-primers li atisa ho kopanngoa ka cartridge ea reagent hantle. Ka kopo etsa bonnete ba hore setsi sa ho qetela sa "conconcentration primer" bakeng sa seliba ke sa ntlha e 'ngoe le e 'ngoe (mohlala, 0.3M ea Index 1 le Index 2 bakeng sa kakaretso ea 0.6M.https://knowledge.illumina.com/library-preparation/general/library-preparation-general-reference_material-list/000001542>

Ka tlase ke

iSeq100
Ka lebaka la khaho ea khatriji ea iSeq100, ha ho khonehe ho kenya le ho sebelisa li-primers tsa tloaelo.

MiniSeq 

NextSeq 

NextSeq 

* Ha ho na boikhethelo bakeng sa setlankana sa tloaelo sa Index 2 (i5) kaha template e sebelisa sehlomathiso sa P5 se kaholimo ho sele ea phallo.

HiSeq 1000/2000 - 1500/2300 

* Ha ho na boikhethelo bakeng sa setlankana sa tloaelo sa Index 2 (i5) kaha template e sebelisa sehlomathiso sa P5 se kaholimo ho sele ea phallo.

HiSeq 3000/4000 

NovaSeq v1.0 Consumables

Kit phetolelo	Illumina Primer (lebitso)	Cartridge Boemo	Kakaretso Bolumo (mL)	Tloaelo se qalang ho qetela ho tsepamisa maikutlo (µM)
SP100/200/300/500 lipotoloho	Bala 1 (BP10)	24	4	0.3
	Index 1 (i7) (BP14)*	23	5	0.3
	Bala 2 (BP11)	13	2	0.3
S1 le S2 100/200/300 lipotoloho	Bala 1 (BP10)	24	4	0.3
	Index 1 (i7) (BP14)*	23	5	0.3
	Bala 2 (BP11)	13	2	0.3
S4200/300 lipotoloho	Bala 1 (BP10)	24	7.3	0.3
	Index 1 (i7) (BP14)*	23	5	0.3
	Bala 2 (BP11)	13	3.5	0.3

* Ha ho na boikhethelo bakeng sa setlankana sa tloaelo sa Index 2 (i5) kaha template e sebelisa sehlomathiso sa P5 se kaholimo ho sele ea phallo.

 

NovaSeq v1.5 Consumables

Kit phetolelo	Illumina Primer (lebitso)	Cartridge Boemo	Kakaretso Bolumo (mL)	Tloaelo se qalang ho qetela ho tsepamisa maikutlo (µM)
SP100/200/300/500 lipotoloho	Bala 1 (VP10)	24	4	0.3
	Index 1 (i7) le Index 2 (i5) (VP14)	23	5	0.3
	Bala 2 (VP11)	13	2	0.3
S1 le S2 100/200/300 lipotoloho	Bala 1 (VP10)	24	4	0.3
	Index 1 (i7) le Index 2 (i5) (VP14)	23	5	0.3
	Bala 2 (VP11)	13	2	0.3
S4200/300 lipotoloho	Bala 1 (VP10)	24	7.3	0.3
	Index 1 (i7) le Index 2 (i5) (VP14)	23	5	0.3
	Bala 2 (VP11)	13	3.5	0.3

NovaSeq X/X Plus 

Ho theha TELL-Seq™ Run ho NextSeq™ 1000/2000 System

The NextSeq™ 1000/2000 reagent cartridge e na le liliba tse peli tsa tloaelo (1 le 2, ka bobeli ha li na letho) tse lokelang ho sebelisoa ha laeborari e hloka bonyane primer e le 'ngoe. E ts'ehetsa ho fihla ho li-primers tse peli tsa tloaelo (letamo) bakeng sa seliba se seng le se seng sa moetlo. Ho bohlokoa hore Read primer le Index primer li ka lilibeng tse fapaneng. Lilaebrari tsa TELL-Seq™ li hloka li-primers tsa tloaelo bakeng sa ho bala hohle (bala 1, bala 2, index 1 le index 2).

Ha lilaebrari tsa TELL-Seq™ li hlophisoa ka tatellano

• Kopanya TELL-Seq™ bala 1 'me u bale li-primers tse 2: sebelisa HT1 ho hlapolla motsoako o mong le o mong oa ho bala oa ho bala ho hlahisa 600 µl ho 0.3 µM ea ho qetela, ke hore, 1.8 µl ea 100 µM TELL-Seq™ bala 1 primer le 1.8 100l µM TELL-Seq™ bala 2 primer ho 597µl ​​HT1. E kenye ka har'a sesebelisoa sa tloaelo 1.
• Kopanya TELL-Seq™ index 1 le index 2 primers: sebelisa HT1 ho hlapolla motsoako o mong le o mong oa li-primer index ho hlahisa 600 µl ho 0.6 µM ea ho qetela, ke hore, 3.6 µl ea 100 µM TELL-Seq™ index 1 primer le 3.6l µM TELL-Seq™ index 100 primer ho 2µl HT593. E kenye ka har'a seliba sa tloaelo 1.

Khetha primer e nepahetseng hantle ha u theha tatellano ea tatellano ka tsela e latelang:

• Bala 1: Tloaelo
• 1 Index
• 1: Tloaelo
• 2 Index 2: Tloaelo
• 2 Bala 2: Tloaelo 1

Ha lilaebrari tsa PhiX li sebelisoa le lilaebrari tsa TELL-Seq™ bakeng sa tatellano ea nako

• Fumana NextSeq 1000/2000 Read Primer Kit (Catalog# 20046117)
• Kopanya TELL-Seq™ bala 1 'me u bale li-primers tse 2 ho HP21: Eketsa motsoako o mong le o mong o tloaelehileng oa ho bala ho 600 µl HP21 bakeng sa 0.3 µM ea ho qetela, ke hore, 1.8 µl ea 100 µM TELL-Seq™ bala 1 primer le 1.8l µM TELL-Seq™ bala 100 primer ho 2µl ​​HP597. E kenye ka har'a sesebelisoa sa tloaelo 21.
• Kopanya TELL-Seq™ index 1 le index 2 primers: sebelisa HT1 ho hlapolla motsoako o mong le o mong oa li-primer index ho hlahisa 600 µl ho 0.6 µM ea ho qetela, ke hore, 3.6 µl ea 100 µM TELL-Seq™ index 1 primer le 3.6l µM TELL-Seq™ index 100 primer ho 2µl HT593. E kenye ka har'a seliba sa tloaelo 1.

Khetha primer e nepahetseng hantle ha u theha tatellano ea tatellano ka tsela e latelang:

• Bala 1: Tloaelo
• 1 Index 1: Tloaelo
• 2 Index 2: Tloaelo
• 2 Bala 2: Tloaelo 1

Ha lilaebrari tsa Illumina tse peli tsa index li hlophisoa le lilaebrari tsa TELL-Seq™ hammoho ka lebelo.

• Fumana NextSeq 1000/2000 Read & Index Primer Kit (Catalog# 20046115)
• Kopanya TELL-Seq™ bala 1 'me u bale li-primers tse 2 ho HP21: Eketsa motsoako o mong le o mong o tloaelehileng oa ho bala ho 600 µl HP21 bakeng sa 0.3 µM ea ho qetela, ke hore, 1.8 µl ea 100 µM TELL-Seq™ bala 1 primer le 1.8l µM TELL-Seq™ bala 100 primer ho 2µl ​​HP597. E kenye ka har'a sesebelisoa sa tloaelo 21.
• Kopanya TELL-Seq™ index 1 le index 2 primer ho BP14: Eketsa motsoako o mong le o mong oa index oa tloaeleha ho 600 µl BP14 bakeng sa 0.6 µM ea ho qetela, ke hore, 3.6 µl ea 100 µM TELL-Seq™ index 1 primer le 3.6l µM TELL-Seq™ index 100 primer ho 2µl BP593. E kenye ka har'a seliba sa tloaelo 14.
• Khetha primer e nepahetseng hantle ha u theha tatellano ea tatellano ka tsela e latelang:
• Bala 1: Tloaelo 1
  Index 1: Tloaelo
• 2 Index 2: Tloaelo
• 2 Bala 2: Tloaelo 1

Lipotso Tse Botsoang Hangata

• P: Ho etsahala'ng haeba tatellano ea Index 1 e sa hlophisoa ka ho feletseng?
  A: Ho hloleha ho tatellano ea Index 1 ka botlalo ho ka lebisa liphosong tsa tlhahlobo ea data e tlase le tlhaloso. Ho bohlokoa ho netefatsa tatellano e felletseng ea Index 1 bakeng sa liphetho tse nepahetseng.
• P: Na ke hloka li-primer tsa tatellano tsa tloaelo bakeng sa litsamaiso tsohle tsa Illumina?
  A: E, li-primers tsa tatellano ea tloaelo lia hlokahala bakeng sa ho hlophisa lilaebrari tsa TELL-Seq TM lits'ebetsong tsohle tsa Illumina ntle le iSeq 100. TheN li-primers li fanoa ho TELL-Seq TM Illumina Sequencing Primer Kit.

Litokomane / Lisebelisoa

tatellano ea UNIVERSAL 100018-USG TELL Seq Library Sequencing [pdf] Bukana ea Mosebelisi 100018-USG TELL Seq Library Sequencing, 100018-USG, TELL Seq Library Sequencing, Seq Library Sequencing, Sequencing Library, Sequencing

Litšupiso

• Bukana ea Mosebelisi