SI -sachin · 2010-06-03 · Title: Microsoft Word - SI_-sachin.doc Author: merisonb Created Date: 6/3/2010 2:35:13 PM

1

Amino/guanidino- functionalized N-(pyrrolidinyl-2-ethyl)glycine- based pet-PNA: Design, synthesis and binding with DNA/RNA Sachin S. Gokhale, Vaijayanti A. Kumar*

Division Of Organic Chemistry, National Chemical Laboratory, Dr. Homi Bhabha Road, Pune-411008, India Email: [email protected]

Contents

Page -2-15 1H, 13C, mass spectra for Compounds 2-11.

Page-16-32 Table of HPLC data and MALDI-TOF data and HPLC chromatograms and

MALDI-TOF spectra for sequences PNA-1 to Gu-petPNA-3(t)

Page-33-36 UV-melting data and UV melting curves

Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is (c) The Royal Society of Chemistry 2010

2

160 150 140 130 120 110 100 90 80 70 60 50 40 30 20

Chloroform-d

21.8

822

.51

23.3

028

.07

30.0

830

.95

46.3

846

.78

53.4

0

76.7

477

.00

77.2

479

.53

79.9

3

117.

68

153.

5315

4.16

Fig. 1:1H NMR spectra for compound 2

Fig. 2:13C NMR spectra for compound 2

NBoc

CN

2

NBoc

CN

2

7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

9.352.722.11 1.351.211.00

Chloroform-d

1.42

1.77

1.85

1.89

1.99

2.01

2.12

2.14

2.54

2.61

2.71

2.73

2.77

3.35

3.38

3.94

4.05

7.25


3

Fig. 3: Mass spectra for compound 2


NBoc

CN

2

NBoc

N

EtO

Cl

O

O

3

7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

9.006.15 5.433.911.24

TMSCDCl3

0.00

1.24

1.26

1.27

1.28

1.30

1.31

1.34

1.47

1.66

1.77

1.86

1.90

1.99

3.33

3.44

3.79

4.01

4.07

4.14

4.21

4.25

7.27


4



NBoc

N

EtO

Cl

O

O

3

NBoc

N

EtO

Cl

O

O

3

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20

Chloroform-d

14.0

1

28.4

030

.83

33.7

0

40.6

941

.11

46.1

246

.91

47.7

449

.49

54.7

454

.96

61.2

261

.81

76.3

777

.00

77.6

479

.35

154.

51

166.

6616

8.62


5



NBoc

N

EtO

T

O

O

4

9 8 7 6 5 4 3 2 1 0

10.674.414.30 3.711.391.131.000.97

TMS

Chloroform-d

0.00

1.23

1.26

1.32

1.46

1.85

1.91

3.293.34

3.403.43

3.73

3.85

4.084.

174.20

4.27

4.444.63

5.31

7.03

7.27

8.77

NBoc

N

EtO

T

O

O

4

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10

Chloroform-d

12.1

613

.94

28.3

329

.92

30.6

933

.46

45.9

146

.48

47.5

147

.96

49.0

954

.71

61.2

861

.93

76.3

677

.00

77.6

379

.33

110.

3011

0.49

140.

97

151.

0815

4.46

164.

4716

6.76

166.

9916

8.77


6



NBoc

N

EtO

T

O

O

4

5

NBoc

N

HO

T

O

O

7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

9.987.804.262.222.12 1.281.00

1.41

1.77

2.51

2.52

2.52

2.53

2.54

3.23

3.26

3.30

3.35

3.62

3.72

3.96

4.06

4.484.

63

7.31

7.43


7



5

NBoc

N

HO

T

O

O

5

NBoc

N

HO

T

O

O

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10

DMSO-d6

12.1

4

28.3

930

.54

32.7

535

.00

38.4

438

.86

39.2

839

.70

40.1

140

.53

40.9

546

.17

47.7

948

.86

54.8

655

.09

78.6

9

108.

26

142.

59

151.

2815

3.84

164.

7216

7.39

170.

6817

1.14


8



N

TBSO

Boc

CN

7

7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

10.239.98 6.541.69 1.39 1.281.141.09 0.70

TMSChloroform-d

0.00

0.07

0.86

1.46

1.94

2.03

2.11

2.18

2.62

2.70

2.72

3.03

3.06

3.11

3.42

4.12

4.39

7.27

N

TBSO

Boc

CN

7

160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0

Chloroform-d

-5.0

2

17.7

5

25.5

228

.25

39.8

540

.87

52.5

255

.27

55.6

9

69.2

069

.68

76.3

677

.00

77.6

479

.98

117.

67

155.

02


9



N

TBSO

BocN

EtO

Cl

O

O

8

7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

9.51 9.007.37 6.343.78 2.35

TMSChloroform-d

0.06

0.85

0.86

1.24

1.27

1.30

1.46

1.56

1.65

1.76

1.98

2.14

3.25

3.33

3.43

3.46

4.00

4.07

4.17

4.21

4.33

7.27


10



N

TBSO

BocN

EtO

Cl

O

O

8

N

TBSO

BocN

EtO

Cl

O

O

8

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0

Chloroform-d

-4.9

4

14.0

217

.82

25.5

828

.34

29.5

933

.78

40.7

141

.10

46.5

847

.73

53.5

754

.97

61.3

0

70.0

276

.36

77.0

077

.64

79.5

5

155.

1215

5.32

155.

36

166.

69


11



9

N

TBSO

BocN

EtO

T

O

O

9 8 7 6 5 4 3 2 1 0 -1

9.82 6.045.24 4.223.96 3.061.131.000.73

TMSCDCl3

0.00

0.04

0.06

0.85

1.26

1.45

1.66

1.91

2.04

3.25

3.26

3.28

3.33

3.41

3.86

3.98

4.17

4.19

4.24

4.33

4.43

4.62

7.03

7.27

8.64

9

N

TBSO

BocN

EtO

T

O

O

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0

Chloroform-d

-4.9

5

12.2

114

.01

17.8

0

25.5

828

.34

29.5

733

.91

40.7

045

.75

47.5

248

.03

53.6

754

.96

61.3

462

.03

70.0

776

.37

77.0

077

.64

79.6

1

110.

4211

0.60

140.

9414

1.09

151.

1115

5.10

164.

3316

6.78

167.

0016

8.78


12



9

N

TBSO

BocN

EtO

T

O

O

N

N3

BocN

EtO

T

O

O

10

9 8 7 6 5 4 3 2 1 0

10.035.82 3.483.032.232.20 1.461.000.88

TMSChloroform-d

0.00

1.24

1.25

1.27

1.29

1.46

1.74

1.83

1.92

1.93

2.11

2.24

2.30

2.35

3.26

3.35

3.39

3.46

3.74

4.10

4.17

4.21

4.32

4.48

4.64

7.03

7.27

8.51


13



N

N3

BocN

EtO

T

O

O

10

N

N3

BocN

EtO

T

O

O

10

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0

12.3

114

.09

22.6

628

.42

29.3

329

.66

30.1

831

.42

31.9

033

.71

46.0

047

.70

47.8

751

.78

54.3

459

.42

61.4

562

.16

76.4

677

.10

77.7

380

.40

110.

57

141.

0514

1.12

151.

1815

4.24

164.

4116

6.96

167.

2916

8.83


14



11

N

FmocHN

BocN

HO

T

O

O

8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0

9.465.024.79 3.002.802.66 2.411.90 1.59

0.87

1.36

1.41

1.54

1.76

2.36

2.51

2.52

2.53

2.61

2.69

2.87

3.02

3.16

3.35

3.62

3.75

3.92

4.23

4.26

4.31

4.33

4.36

4.46

4.49

4.63

7.31

7.35

7.38

7.43

7.47

7.56

7.60

7.69

7.72

7.89

7.92

11

N

FmocHN

BocN

HO

T

O

O

170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20

12.3

1

28.5

729

.42

31.6

1

44.4

547

.19

48.1

350

.83

54.1

5

65.8

5

79.0

4

108.

4211

0.16

120.

5612

1.83

125.

6112

7.52

128.

0712

9.39

137.

9013

9.89

141.

2114

2.74

144.

3515

1.52

153.

8915

6.25

164.

8916

7.57

171.

41


15


11

N

FmocHN

BocN

HO

T

O

O


16

Oligomer Synthesis: The PNA oligomers were synthesized by standard Boc-solid phase peptide

strategy. Purification of all the PNA oligomers was carried out by Varian dual pump PROSTAR

model No.210 HPLC on RP-C18 column with Water: acetonitrile-0.1% TFA system. The

gradient used for HPLC analysis is (A: 95% Water: 5% acetonitrile+0.1% TFA, B: 50% Water:

5% acetonitrile-0.1% TFA ) 0-20 min 100% B. PNA oligomers were characterized by MALDI-

TOF mass spectrometry by using Voyager-DE-STR (Applied Biosystems) MALDI-TOF. A

nitrogen laser (337 nm) was used for desorption and ionization. Spectra were acquired in linear

mode. The matrix used for analysis was CHCA (α-cyano hydroxyl cinnamic acid)

Table.1 PNA Oligomers synthesized their HPLC tR and MALDI-TOF mass characterization.

Entry Sequence HPLC tR (min)

Mass calcd/obsd

PNA-1 H-TTTTTTTT-Lys-NH2( C94H127N35O33) 8.90 2273.93/2274.91 H-petPNA-1(s) Am-petPNA-1(s) Gu-petPNA-1(s)

H-TTTTtRTTT-Lys-NH2 R = H( C98H134N35O33) R= NH2(C98H135N36O33) R= NH-C(= NH)NH2 (C99H137N38O33)

9.21

14.16* 14.58*

2328.98/2329.1 2343.99/2343.34 2386.02/2382.33

H-petPNA-1(s*) Am-petPNA-1(s*) Gu-petPNA-1(s*)

H-TTTTTTTtR-Lys-NH2 R = H( C98H134N35O33) R= NH2(C98H135N36O33) R= NH-C(= NH)NH2 (C99H137N38O33)

9.48 8.48 8.66

2328.98/2329.20 2343.99/2366.13(M+Na+) 2386.02/2386.57

PNA-2 H-TTACCTCAGT-Lys-NH2 (C113H149N55O33)

9.04 2804.16/2827.0(M+Na+)

H-petPNA-2(s) H-TTACCtRCAGT-Lys-NH2 R=H(C117H156N55O33)

9.30 2859.22/2859.38

H-petPNA-2(d) Am-petPNA-2(d) Gu-petPNA-2(d)

H-TTACCtRCAGtR-Lys-NH2 R=H (C121H163N55O33) R= NH2 (C121H165N57O33) R= NH-C(=NH)NH2 (C123H169N61O33)

9.60

12.98* 14.12*

2914.27/2910.43 2944.29/2940.6 3028.34/3024.5

PNA-3 H-CTTCTTCCTT-Lys-NH2 (C112H151N47O37)

11.19 2746.13/2741.15

Am-petPNA-3(t) Gu-petPNA-3(t)

H-CTtRCTtRCCtRtR-Lys-NH2 R= NH2 (C128H183N51O37) R= NH-C(= NH) NH2 (C132H191N59O37)

13.47* 14.64*

3026.40/3034.50 3194.35/3201.40

* The gradient use for HPLC analysis is (A: 95% Water: 5% acetonitrile-0.1% TFA, B: 50%

Water: 50% acetonitrile-0.1% TFA) 0-20 min- 0%-50%B, 20-30min-50%-100% B


17

Fig. 28: HPLC chromatogram for PNA-1

Fig. 29: HPLC chromatogram for H-petPNA-1(s)

PNA-1

H-TTTTTTTT-Lys-NH2

H-petPNA-1(s)

H-TTTtHTTTT-Lys-NH2


18

Fig. 30: HPLC chromatogram for Am-petPNA-1(s)

Fig. 31: HPLC chromatogram for Gu-petPNA-1(s)

Fig. 32: HPLC chromatogram for Am-petPNA-1(s) +Gu-petPNA-1(s)

Name Retention Time Area % Area Height Int Type 1 13.625 21779 1.55 2293 BV 2 14.163 1379306 98.45 44243 VB

Name Retention Time Area % Area Height Int Type 1 1.494 1969 0.05 288 BB 2 14.581 3658315 99.95 132240 BB

Name Retention Time Area % Area Height Int Type 1 2.115 26271 1.23 275 BB 2 13.655 24430 1.14 2094 BV 3 14.155 765374 35.71 36101 VV 4 14.597 1327210 61.92 48743 VB

13.6

2514

.163

AU

0.000

0.010

0.020

0.030

0.040

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Am-petPNA-1(s)

H-TTTtNH2TTTT-Lys-1.

494

14.5

81

AU

0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Gu-petPNA-1(s)

H-TTTtGTTTT-Lys-NH2

2.11

5

13.6

5514

.155

14.5

97

AU

0.000

0.010

0.020

0.030

0.040

0.050

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Gu-petPNA-1(s) + Am-petPNA-1(s)


19

Fig. 33: HPLC chromatogram for H-petPNA-1(s*)

Fig. 34: HPLC chromatogram for Am-petPNA-1(s*)

H-petPNA-1(s*)

H-TTTTTTTtH-Lys-NH2

Am-petPNA-1(s*)

H-TTTTTTT tNH2-Lys-NH2


20

Fig. 35: HPLC chromatogram for Gu-petPNA-1(s*)


Gu-petPNA-1(s*)

H-TTTTTTTtG-Lys-NH2

PNA-2

H-TTACCTCAGT-Lys-NH2


21

Fig. 37: HPLC chromatogram for H-petPNA-2(s)

Fig. 38: HPLC chromatogram for H-pet PNA-2(d)

H-petPNA-2(S)

H-TTACCtHCAGT-Lys-NH2

H-petPNA-2(d)

H-TTACCtHCAGtH-Lys-NH2


22

Fig. 39: HPLC chromatogram for Am-petPNA-2(d)

Fig. 40: HPLC chromatogram for Gu-petPNA-2(d)

Fig. 41: HPLC chromatogram for Am-petPNA-2(d) +Gu-petPNA-2(d)

Name Retention Time Area % Area Height Int Type 1 12.988 180969 100.00 7851 BB



13.2

57

14.1

51AU

0.000

0.005

0.010

0.015

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00

Am-petPNA-2(d) + Gu-petPNA-2(d)

13.2

35 14.1

26

AU

0.000

0.005

0.010

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00

Gu-petPNA-2(d)

H-TTACCtGCAGtG-Lys-NH2

12.9

88

AU

0.000

0.002

0.004

0.006

0.008

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00

Am-petPNA-2(d)

H-TTACCtNH2CAGtNH

2-Lys-


23


Fig. 43: HPLC chromatogram for Am-petPNA-3(t)

Fig. 44: HPLC chromatogram for Gu-petPNA-3(t)

Name Retention Time Area % Area Height Int Type 1 1.451 56443 0.83 18197 BV 2 1.519 153164 2.24 35130 VB 3 10.120 78711 1.15 4946 VV 4 10.477 71552 1.05 6835 VV 5 11.198 6471200 94.73 305003 VB

Name Retention Time Area % Area Height Int Type 1 1.399 1277 0.27 165 BB 2 13.476 478492 99.73 11055 BB

Name Retention Time Area % Area Height Int Type 1 1.365 700 0.12 105 BB 2 3.191 5183 0.90 70 BB 3 13.745 5278 0.91 375 BV 4 14.649 565951 98.07 15153 VB

1.45

11.

519

10.1

2010

.477

11.1

98

AU

0.00

0.20

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

PNA-3 H-CTTCTTCCTT-Lys-

1.39

9

13.4

76

AU

0.000

0.002

0.004

0.006

0.008

0.010

0.012

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Am-petPNA-3(t)

H-CTtNH2CTtNH

2CCtNH2tNH

2-Lys-NH2

1.36

5

3.19

1

13.7

45

14.6

49

AU

0.000

0.005

0.010

0.015

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Gu-petPNA-3(t)

H-CTtGCTGCCtGtG-Lys-NH2


24

Fig. 45: HPLC chromatogram for Am-petPNA-3(t) + Gu-petPNA-3(t)

Name Retention Time Area % Area Height Int Type 1 3.174 12032 0.78 176 BB 2 13.325 739822 47.81 16480 BV 3 14.618 795616 51.41 21296 VB

3.17

4

13.3

25

14.6

18

AU

0.000

0.005

0.010

0.015

0.020

Minutes0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

Am-petPNA-3(t) + Gu-petPNA-3(t)


25

Fig. 46: MALDI-TOF spectra for PNA-1

Fig. 47: MALDI-TOF spectra for H-petPNA-1(s)

PNA-1 H-TTTTTTTT-Lys-NH2

Mass Calcd. 2273.93

Mass Obsd. 2274.91

H-petPNA-1(s) H-TTTTtHTTT-Lys-NH2

Mass Calcd. 2329.98

Mass Obsd. 2329.18


26

Fig. 48: MALDI-TOF spectra for Am-petPNA-1(s)

Fig. 49: MALDI-TOF spectra for Gu-petPNA-1(s)

Am-petPNA-1(s)

H-TTTTtNH2TTT-Lys-NH2

Mass Calcd. 2343.99

Mass Obsd. 2343.31

Gu-petPNA-1(s)

H-TTTTtGTTT-Lys-NH2

Mass Calcd. 2386.02

Mass Obsd. 2382.33


27

Fig. 50: MALDI-TOF spectra for H-petPNA-1(s*)

Fig. 51: MALDI-TOF spectra for Am-petPNA-1(s*)

H-petPNA-1(s*)

H-TTTTTTTtH-Lys-NH2

Mass Calcd. 2328.98

Am-petPNA-1(s*)

H-TTTTTTTtNH2-Lys-NH2

Mass Calcd. 2343.99

Mass Obsd. 2344.04 (M+ Na+)


28

Fig. 52: MALDI-TOF spectra for Gu-petPNA-1(s*)


Gu-petPNA-1(s*)

H-TTTTTTTtG-Lys-NH2

Mass Calcd. 2386.02

Mass Obsd. 2386.57

PNA-2

H-TTACCTCAGT-Lys-NH2

Mass Calcd 2804.16

Mass Obsd. 2827.0 (M+Na+)


29

Fig. 54: MALDI-TOF spectra for H-petPNA-2(s)

Fig. 55: MALDI-TOF spectra for H-petPNA-2(d)

H-petPNA-2(d)

H-TTACCtHCAGtH-Lys-NH2

Mass Calcd 2914.27

Mass Obsd. 2910.43

H-petPNA-2(s)

H-TTACCtHCAGT-Lys-NH2

Mass Calcd 2859.22

Mass Obsd. 2859.38


30

Fig. 56: MALDI-TOF spectra for Am-petPNA-2(d)

Fig. 57: MALDI-TOF spectra for Gu-petPNA-2(d)

Am-petPNA-2(d)

H-TTACCtNH2CAGtNH

2-Lys-NH2

Mass Calcd 2944.21

Mass Obsd. 2940.63

Gu-petPNA-2(d)

H-TTACCtGCAGtG-Lys-NH2

Mass Calcd 3028.34

Mass Obsd. 3024.52


31


Fig. 59: MALDI-TOF spectra for Am-petPNA-3(t)

PNA- 3

H-CTTCTTCCTT-Lys-NH2

Mass Calcd. 2746.13

Mass Obsd. 2741.15

Am-petPNA-3(t)

H-CTtNH2CTtNH

2CCtNH2tNH

2-Lys-NH2

Mass Calcd 3026.40

Mass Obsd. 3034.50


32

Fig. 60: MALDI-TOF spectra for Gu-petPNA-3(t)

Gu-petPNA-3(t)

H-CTtGCTtGCCtGtG-Lys-NH2

Mass Calcd 3194.35

Mass Obsd. 3201.40


33

UV-Tm measurements:

Entry Sequence DNA-1 (DNA-3)

RNA-1 (RNA-3)

PNA-1. H-TTTTTTTT-Lys-NH2 41.6 (30.0) 41.2 (29.8) H-petPNA-1(s) Am-petPNA-1(s) Gu-petPNA-1(s)

H-TTTTtRTTT-Lys-NH2

R= H

R= NH2

R= NH-C(= NH) NH2

27.2 (nd) 40.7 (19.4) 36.01 (21.4)

37.4(16.5) 36.8(18.2) 36.4(18.9)

DNA-1: 5’GCAAAAAAACG 3’ RNA-1: 5’GCAAAAAAACG 3’ DNA-3: 5’GCAAATAAACG 3’ RNA-3: 5’GCAAAUAAACG

Fig. 61: A UV melting Curves of PNA-1-Gu-petPNA-1(s) with DNA-1 and B corresponding derivative graphs

Fig. 62: A UV melting Curves of PNA-1-Gu-petPNA-1(s) with RNA-1 and B corresponding derivative graphs


34


RNA-1 (RNA-3)

PNA-1. H-TTTTTTTT-Lys-NH2 41.6 (30.0) 41.2 (29.8) H-petPNA-1(s*) Am-petPNA-1(s*) Gu-petPNA-1(s*)

H-TTTTTTTtR-Lys-NH2

R= H

R= NH2

R= NH-C(= NH) NH2

26.7 (nd) 45.7 (26.8) 51.6 (32.6)

48.7(20.9) 49.9(28.5) 58.7(31.8)

DNA-1: 5’GCAAAAAAACG 3’ RNA-1: 5’GCAAAAAAACG 3’ DNA-3: 5’GCAAATAAACG 3’ RNA-3: 5’GCAAAUAAACG

Fig. 63: A UV melting Curves of PNA-1-Gu-petPNA-1(s*) with DNA-1 and B corresponding derivative graphs Fig. 64: A UV melting Curves of PNA-1-Gu-petPNA-1(s*) with RNA-1 and B corresponding derivative graphs


35

DNA-2: 5’ ACTGAGGTAA3’ RNA-2: 5’ UGUAACUGAGGUAAGAGG 3’ DNA-4: 5’ ACTGTGGTAA3’ RNA-4: 5’ UGUAACUGCGGUAAGAGG DNA-5: 5’ AATGGAGTCA 3’

Fig. 65: A UV melting Curves of PNA-2-Gu-petPNA-2(d) with DNA-2 and B corresponding derivative graphs

Fig. 66: A UV melting Curves of PNA-2-Gu-petPNA-2(d) with RNA-2 and B corresponding derivative graphs


RNA-2 (RNA-4)

pDNA-5 (∆ Tm= ap-p)

PNA-2 H-TTACCTCAGT-Lys-NH2 54.2 ( 47.5) 54.5(46.5) 53.9 (+0.3) H-pet-PNA-2(s) H-TTACCtCAGT-Lys-NH2

R= H

57.6 (51.2)

52.6 (38.8)

55.4 (+1.8)

H-pet-PNA-2(d) Am-pet-PNA-2(d) Gu-pet-PNA-2(d)

H-TTACCtRCAGtR-Lys-NH2 R= H R= NH2

R= NH-C(= NH) NH2

63.2 (50.8) 66.9 (58.1) 68.9 ( 60.0)

48.1 (34.5) 52.1 (43.8) 54.2 (46.1)

59.3 (+3.9) 59.5 (+7.4) 59.9 (+9.0)


36


RNA-1 (RNA-3)

PNA-3 H-CTTCTTCCTT-Lys-NH2 52.0 (42.6) 55.6 (45.2) Am-petPNA-3(t) Gu-petPNA-3(t)

H- CTtRCTtRCCtRtR-Lys-NH2

R= NH2

R= NH-C(= NH) NH2

63.7(49.9) 64.8 (54.5)

65.5(57.6) 68.3(59.2)

DNA -6: 5’AAGGAAGAAG 3’ RNA-6: 5’ AAGGAAGAAG 3’ DNA 7: 5’ AAGGTAGAAG 3’ RNA 7: AAGGUAGAAG 3’

Fig. 67: A UV melting Curves of PNA-3-Gu-petPNA-3(t) with DNA-6 and B corresponding derivative graphs

Fig. 68: A UV melting Curves of PNA-3-Gu-petPNA-3(t) with RNA-6 and B corresponding derivative graphs


Documents

SI -sachin · 2010-06-03 · Title: Microsoft Word - SI_-sachin.doc Author: merisonb Created Date: 6/3/2010 2:35:13 PM