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o<br />

Ó^,<br />

MATERIALES Y MÉTODOS<br />

Tabla 3.3. Diseño de los cebadores para las reacciones de amplificación y selección de las<br />

enzimas utilizadas en la comprobación de cada producto obtenido Tm: temperatura de<br />

melting, L.P.: Longitud del producto de PCR; L: longitud del cebador. (continuación).<br />

Nombre<br />

del<br />

Cebador<br />

Posición del<br />

Cebador<br />

Inicio final<br />

SECUENCIA (ó^ (nt) ^^ Enzima<br />

Longitud<br />

Fragmentos<br />

(pb)<br />

,<br />

.<br />

,G31^10 3^1^1Ot5' ' `313 ŜÓ9^ 3^13'Ŝ28; GCTCTGGCCGTCGTTGGAAC<br />

" (3,5 20 65'Ŝ PvúI1 _. ... 376^278 . .:<br />

3"110 `,3' ^314^163, 31'41x42 GACACGGACCAAACTTCGCC:AT_ 62,1. 22 : _ '<br />

G3113 3113-5' 315037 315058 CCAAGGAGGCCTGAATGGAACG 64,0 22 588 B 111 356+231<br />

^'G3116<br />

3113-3'<br />

YGL98 183<br />

315624 315603 ACCGGGCAGAGGTGGCAATAAA<br />

_<br />

.<br />

62,1<br />

317522 3175"42 ^GCAAACAAAACATGACGCCCA 57;9<br />

22<br />

2i<br />

, ..<br />

336<br />

.<br />

C1iiI 253+82<br />

YGL98-498 ' •31^7857 3^17837; GĈTCCTTGCTTCAAACTGCCC 61;8 21 ^ "<br />

G3146 3146-5' 325369 325383 GCCCCCAATCCAGGT 47 4 15 333 H aI 202+131<br />

^Ĝ3168:<br />

3146-3'<br />

. ..<br />

3.,1`68'=5'<br />

.<br />

3_168;'-'3'<br />

325701<br />

..<br />

325687<br />

_<br />

334895<br />

..<br />

334915;<br />

33550.4; 335483<br />

_<br />

CGGGCTGCGAAGAAG<br />

CGAAAGAAGCCGGAITGGGGG<br />

.<br />

TGTTfCĈTCTGGCACTGCCTCG<br />

47,4 15<br />

61=8 2^1<br />

_<br />

64;0, ' 22<br />

_ _<br />

61U:<br />

"<br />

,.<br />

Cl^il<br />

^<br />

364^213+40,;:<br />

G3175 3175-5' 342661 342646 CCGTCCATGAATCGAA 49,2 16 479 Dral 336+142<br />

,G31^79<br />

3175-3'<br />

..<br />

3^1:79.=5'<br />

342183 342197<br />

34370..7 343722;<br />

CGGAACAGACCCCAA<br />

TTGG^GTGAATCGGCGÁ<br />

50,6<br />

47,8"<br />

15<br />

1'6 374 .- HiiidLll<br />

^" 20.:4,+170<br />

3179=3'^ 344080 344065' GGCTITTGCACTTCCG ,46;6 16.,, " ^ ' ^ _ . ^ -<br />

G3189 3189-5' 346762 346747 AAGGGTTTGGTCCAGA 41,5 16 341 Ncol 267+73<br />

.. ...<br />

: G3f93<br />

3189-3'<br />

.<br />

3:193=5'<br />

31:93:-3'<br />

346422<br />

. ...<br />

346436<br />

. ..<br />

35,O1Q6<br />

.<br />

35Q12.1;<br />

.<br />

350418 3504Q3..<br />

.<br />

CTTCGGTTGACGCAA<br />

. .<br />

AAĈCCAGCTCAÁGCCrT<br />

_<br />

CACCAGGCGTT.TCAAA<br />

42,3<br />

.<br />

44;8.<br />

44;0:<br />

15<br />

-<br />

".:<br />

16<br />

..<br />

1^6<br />

-<br />

313<br />

. ,^<br />

Fliñĉl711. 158+'1:30±25<br />

G3210 3210-5' 361452 361437 ACGCCTATTCAAGCCG 51,7 16 488 EcoRl 271+217<br />

.G3406^:<br />

3210-3'<br />

.<br />

3406=5'<br />

360965 360980<br />

: .<br />

3,79Q58 379073;<br />

GGCCAGGTAGCGGTAA<br />

. ..<br />

CCÁGGCGÁGTACCCAA ^<br />

54,3<br />

46 3;<br />

16<br />

".<br />

.16 433.,<br />

" . ._<br />

EĉóRl ,<br />

.<br />

369±;^63 .<br />

3406`3' . 37949;b 3,7.947Ŝ: TACÁCCT'TGCCĈĜGÁG 46,5; 16 " ^ "<br />

G3432 YGL061c5' 389597 389577 CAGGAGGTGGAGCAAGCGATA 61 8 21 551 Alul 272+235+43<br />

'G3435"<br />

YGL061c3'<br />

3435:'S'<br />

389047 389066<br />

...<br />

390456 390471;<br />

CGAGGCAGGAACATGGGTCT<br />

,<br />

= CTCCGG^IGATT,GGGCA<br />

61,4 20<br />

"<br />

49,2^ , lb<br />

.<br />

58;1 Cl^il<br />

.....<br />

443^1^38<br />

3„43Ŝ=3' ; 391Q36 391021; GGCCGGGTTCt^TGAG^ 48,9, "16.,., ^ . - " , -<br />

G3474 3474-5' 401283 401268 TGGGTGCTTGGTTATT 46 6 16 318 H al 190+128<br />

3474-3' 400966 400981 CCCTTI'rATGTTTGCC 46,6 16<br />

Ĝ3483 3483-5'<br />

34,83=3'<br />

402635,<br />

..:<br />

,4U2v65Q;<br />

_.<br />

403055 403U4.1:<<br />

AAĈĈCGGTGGATATT'T<br />

.<br />

C,GGTGGĈAATCITCA<br />

46,6<br />

: .__<br />

47;8<br />

_ 16;: 42'1 - IV"dél .. 2, 50^1^1<br />

,.<br />

.<br />

^<br />

1_5<br />

"<br />

G3489 3489-5' 404038 404053 CCGGCAGTAGAAGGGA 54,3 16 330 Clal 321+9<br />

G3505<br />

3489-3'<br />

^<br />

3,SOS=S'<br />

404367 404352<br />

405776 ,.405791,;.<br />

CGCATCGATATTGGGA<br />

, GCTGC.GĈY^GAÁTGCCT<br />

49,2 16<br />

Ŝ0,0. ^ 16 542 1Í^16qÍ 281^147+114<br />

35Q5.-3' 406317 406302:": CCGAGCĜGTACGTCCÁ ŜU,S T6:^. ..:" . . .<br />

G3517 3517-5' 411580 411595 TTACGTGTGGGGCAAC 44 3 16 329 EcoRl 273+56<br />

3517-3' 411908 411893 CGTTCCAGCGTGTGAT 44,1 16<br />

G3535 3Ŝ35=5' 4,1454'1 4,14556'; CCACGAGGCAGAGG ,A'A , 46,3 1:6 330 ,Yhól 172±T58 _<br />

3535;:3' , 414870 4,1,4855; ._ ,GG.GĜGAC,"CJ,TCTTGGAG 47,^3, 16__ .<br />

G3540 3540-5' 415140 415155 GCCCCTTGATGCATTT 45,6 16 535 Alul 320+215<br />

_,<br />

G3679.<br />

3540-3'<br />

3679.,, Ŝ'<br />

..<br />

3679'3' ^.<br />

415527 415512<br />

45169.4<br />

.:<br />

:4516;7,9.<br />

.<br />

45,1223 451237;<br />

TGAGGATTGGGATGGG<br />

.<br />

AGCTCGGTGAAĜGC^Á<br />

^<br />

- TGATGCGGCGGTTTC<br />

45,6<br />

..<br />

49,0<br />

,4,8,4<br />

16<br />

16..<br />

15<br />

.<br />

472 .<br />

: .<br />

^^ ^<br />

.<br />

, .<br />

ĉiLUl<br />

, `<br />

_..<br />

385±8,7..<br />

G3690 3690-5' 457856 457841 CAGCAGCGGTAGCGAT 47,2 16 499 EcoRl 350+149<br />

G3,719:<br />

3690-3'<br />

37,19,5'<br />

37-19=3'<br />

457358 457373<br />

-<br />

.<br />

4683U.0 468315;;<br />

468739, 468724;<br />

CCAAATTTGCCCTCCA<br />

_ _<br />

.^; GGGTGTTGCGTTTTĜĈ<br />

"<br />

; ACĈAGTAAAGGCGCĈA<br />

46,6<br />

"<br />

51,7<br />

^<br />

5;1,7<br />

16<br />

,..<br />

l'6<br />

16<br />

44Q<br />

,.: ^<br />

_ _ .<br />

Bám'Fll<br />

. -<br />

.<br />

342±98<br />

G3759 3759-5' 490476 490461 GTTCCCCATGCATTCG 46,6 16 510 Hindlll 258+252<br />

3759-3' 489967 489982 ATCTGTCCCCGTGCTG 45 9 16<br />

47

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