-80 freezer in 450 N. Frear
Open space.
Rack A (Dave's rack):
1. DSG Antibodies #1 (Pol II antibody);
2. GAGA factor Ab Box;
3. Autin's transcription factors;
4. DSG antibodies #2;

5. DSG Expression #1;
6. DSG Proteins #1;
7. DSG antibodies #3;
8. GAGA factor;

9. Conaway's general factors;
10. Bacteria Strains #1;
11. Austin's 2nd shipment of factors;
12. Gilmour's TFIID;

13. Price/Greenleaf;
14. RNA polymerase II
15. Competent BL21(DE3)

Missing boxes:
DSG Proteins #2; 

 Rack B
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.Dave Temp: Emanuel's cells: IIB, dTBP, numerous hsp70 promoters from deletion work.
14. Gregory's IIB from Fall 94, column fractions.
15. Gregory's IIB from Fall 94
16. FITC labelled secondaries and Gal4 Ab		 Rack C
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack D
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 
Rack E (Larry's rack)
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack F (Larry's rack plus other stuff)
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack G
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack H
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 
Rack I
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack J
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack K
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 		 Rack L
1.
2.
3.
4.

5.
6.
7.
8.

9.
10.
11.
12.

13.
14. 
15. 
16. 
Bottom row reserved for autoradiograms

DSG Antibodies #1:
Notes: All of these pol II antibodies were prepared from injection done during the Summer of 1992 with one rabbit.

1. Pol II preimmune serum; 5/29/92; Rabbit # 5277
2. Pol II preimmune serum; 5/29/92; Rabbit # 5277
3. Pol II preimmune serum; 5/29/92; Rabbit # 5277; 100ul
4. Pol II preimmune serum; 5/29/92; Rabbit # 5277; 100ul
5. Pol II preimmune serum; 5/29/92; Rabbit # 5277; 100ul
6. Pol II preimmune serum; 5/29/92; Rabbit # 5277; 100ul

7-21. Pol II serum, first bleed; Rabbit # 5277

22-36. Pol II serum, 2nd bleed; Rabbit # 5277

37-81. Pol II serum, final bleed; Rabbit # 5277; inadvertantly labeled 9/93


DSG antibodies #2:
1-12. Pol II serum, final bleed; Rabbit # 5277 (also see DGS antibodies #1)

13. TBP rabbit serum, from Hoey/Tjian, 1992
14. TBP rabbit serum, from Hoey/Tjian, 1992
15. TBP rabbit serum, from Hoey/Tjian, 1992
16. TBP rabbit serum, from Hoey/Tjian, 1992
17. TBP rabbit serum, from Hoey/Tjian, 1992
18. TBP rabbit serum, from Hoey/Tjian, 1992
19. TBP rabbit serum, from Hoey/Tjian, 1992
20. TBP rabbit serum, from Hoey/Tjian, 1992
21. TBP rabbit serum, from Hoey/Tjian, 1992
22. TBP rabbit serum, from Hoey/Tjian, 1992
23. TBP rabbit serum, from Hoey/Tjian, 1992

24. Rabbit preimmune for Hoey/Tjian TBP serum, #9307

25. HSF antibody, 944-3, lyophilized powder from 50 ul, Wu/Tsukiyama, 2/16/95
26. HSF antibody, 944-3, lyophilized powder from 50 ul, Wu/Tsukiyama, 2/16/95
27.
28.
29.

30. TFIIB rabbit serum from Kadonaga, (obtained by P. Emanuel)

31. GAGA rat serum from Ganock/Elgin, Fall 93 (NM)
32. GAGA rat serum from Ganock/Elgin, Fall 93 (NM)
33. GAGA rat serum from Ganock/Elgin, Fall 93 (NM)

34. GAGA rat serum from Ganock/Elgin, Fall 93 (1H)
35. GAGA rat serum from Ganock/Elgin, Fall 93 (1H)
36. GAGA rat serum from Ganock/Elgin, Fall 93 (1H)

37. GAGA rat serum from Ganock/Elgin, Fall 93 (1T)
38. GAGA rat serum from Ganock/Elgin, Fall 93 (1T)
39. GAGA rat serum from Ganock/Elgin, Fall 93 (1T)

40. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (1-H), 50% ethylene glycol
41. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (1-H), 50% ethylene glycol
42. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (1-H), 50% ethylene glycol

43. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (2-H), 50% ethylene glycol
44. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (2-H), 50% ethylene glycol
45. GAGA rat preimmune serum from Ganock/Elgin, Fall 93 (2-H), 50% ethylene glycol

46. GAGA rabbit serum from Wu, Spring 94
47. GAGA rabbit serum from Wu, Spring 94
48. GAGA rabbit serum from Wu, Spring 94
49. GAGA rabbit serum from Wu, lyophilized

50. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
51. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
52. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
53. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
54. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
55. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
56. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
57. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
58. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
59. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media
60. TAFII250 mouse monoclonal, 2B2, Weinzerl/Tjian, 94, culture media

61. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media
62. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media
63. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media
64. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media
65. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media
66. TAFII250 mouse monoclonal, 30H9, Weinzerl/Tjian, 94, culture media - transferred to Baiyong, 7/28/94

67. TAFII150 rabbit serum, 823, Verrijzer/Tjian, 94
68. TAFII150 rabbit serum, 823, Verrijzer/Tjian, 94
69. TAFII150 rabbit serum, 823, Verrijzer/Tjian, 94

70. Drosophila PolII goat IgG fraction from Anro Greenleaf, 63N IgG
71. Drosophila PolII goat IgG fraction from Anro Greenleaf, 63N IgG
72. Drosophila PolII goat IgG fraction from Anro Greenleaf, 63N IgG

73. Drosophila TFIIB rabbit serum from Wampler/Kadonaga
74. Drosophila TFIIB rabbit serum from Wampler/Kadonaga
75. Drosophila TFIIB rabbit serum from Wampler/Kadonaga
76. Drosophila TFIIB rabbit serum from Wampler/Kadonaga
77.
78.
79.
80.
81.


DSG antibodies #3:
Notes: Antiserum in slots 1 - 25 are quite old and were stored many years in the refrigerator.  They should be tested prior to use.

Slots 1-9, Lambda repressor rabbit serum, 7/28/82

Slots 10-23, E. coli RNA polymerase rabbit serum, 3/23/82

24. H4 rabbit serum, (Ra 447c) D. Stollar, 7/18/83
25. H3/H4 rabbit serum, (Ra 372d) D. Stollar, 7/18/83

Slots 26-31, TBP rabbit serum, #5278, Peter Emanuel, 1/28/93 (1ml)

Slots 32-50, TBP rabbit serum, #5317, Peter Emanuel, 5/5/93 (1ml)

Slots 51-64, TAF250 monoclonal, 30H9, hybridoma supernatant from Tjian lab, provided 5/19/00 by P. Verrijzer

Slots 65-78, TAF250 monoclonal, 2B2, hybridoma supernatant from Tjian lab, provided 5/19/00 by P. Verrijzer

79. TAF150 polyclonal made in rabbit #823 in Tjian's lab.  Provided 5/19/00 by P. Verrijzer, designated 1 on the lid and should be good.

80.  TAF150 polyclonal made in rabbit #823 in Tjian's lab.  Provided 5/19/00 by P. Verrijzer, designated 2 on the lid, this is the bleed-out and apparently has high background.

81. TAF150 polyclonal made in rabbit #824 in Tjian's lab.  Provided 5/19/00 by P. Verrijzer, designated 3 on lid, this is a test bleed and should be good.


GAGA factor antibody:  Prepared by Deb Taxman in 1995 against recombinant GAGA factor.  Aliquoted by Yingyun Lui on 8/22/99.

Slots 1-4: pre-immune, 1.3ml/tube, 3/20/95
Slots 5-8: first bleed sera, 1.3ml/tube, 4/17/95 - red label
Slots 9-12: second bleed sera, 1.3ml/tube, 5/3/95 - green label
Slots 13-62: Final bleed sera,  1.3ml/tube, 5/31/95
Slots 71-81: Final bleed sera, 0.1 ml/tube, 5/31/95


DSG Expression #1:
1. BL21 DE3 glycerol stock

2. dTBP expression, pAR3038 dTBP, glycerol stock in BL21 DE3 (Tjian)
3. dTBP expression, pAR3038 dTBP, glycerol stock in BL21 DE3 (Tjian)
4. space for dTBP DNA

5. dTFIIB expression, pET dTFIIB2, glycerol stock in BL21 DE3 (Kadonaga)
6. dTFIIB exoression, pET dTFIIB2, glycerol stock in BL21 DE3 (Kadonaga)
7. pET dTFIIB2, 20 ng/ul

8. pAR3040 56KTFIIE, (human) glycerol stock in TG2 from Tjian via Rick Austin/Biggin
9. pAR3040 34KTFIIE, (human) glycerol stock in TG2 from Tjian via Rick Austin/Biggin
10. pET-RAP74, (human) glycerol stock in unknown strain from Sherm Weissman via Austin
11. pET-RAP30, (human) glycerol stock in unknown strain from Sherm Weissman via Austin

12. Drosophila GAGA factor: pARGAGA, DNA at 0.25 mg/ml from Soeller via Granock/Elgin
13. Drosophila GAGA factor: pETGAGA, DNA at 0.95 mg/ml from Kadonaga via Granock/Elgin
14. Drosophila GAGA factor: pETGAGA, DNA at 0.95 mg/ml from Kadonaga via Granock/Elgin

15. Drosophila heat shock factor: pHSF20, DNA at 1 mg/ml from C. Wu
16. pEZZ18, DNA at 0.5 mg/ml from Pharmacia, for Protein A-like fusions.
17. pEZZ-GAGA (A13), miniprep DNA, putative GAGA fusion by Gilmour (summer 94)
18. pEZZ-GAGA (A15), miniprep DNA, putative GAGA fusion by Gilmour (summer 94)
19. pEZZ-GAGA rev (A5), miniprep DNA, putative GAGA fusion by Gilmour
20. pEZZ-GAGA rev (A16), miniprep DNA, putative GAGA fusion by Gilmour

21.Micrococcal nuclease: pFOG 301, DNA 0.5mg/ml from David Shortle
22. GST/protein A fusion: pAGEX2T, DNA 0.5 mg/ml from Alan Wildeman

23. dHSF from baculovirus, Mary Fernandez/Lis, 9/92, in 20 mM Hepes, 10%glycerol, 0.3 M NaCl, not homogeneous.
24. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
25. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
26. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
27. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
28. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
29. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
30. dHSF from baculovirus, Mary Fernandez/Lis, 9/92
31. dHSF from baculovirus, Mary Fernandez/Lis, 9/92

32-52. dTFIIB, Walters, Emanuel and Sypes, Hydroxyapatite fraction 23, quality unknown, Derived from Kadonaga's pET dTFIIB2.

53-71. dTFIIB, Walters, Emanuel and Sypes, Hydroxyapatite fraction 24, 130 ng/ul in 100 mM HGKEDP, 90% pure but may have a little contaminating endonuclease.

72. Avidin DN, A-3100, Vector Lab, 1 mg/ml, spring 94
73. Protein G- Biotin, Sigma, PB045, 2 mg/ml, spring 94

74-81. dTFIIB, Walters, Emanuel and Sypes, Hydroxyapatite fraction 23, quality unknown


GAGA factor:
1. CT-2 F1 from 2/17/89, 2.5 ul will footprint
2. CT-2 F1 from 2/17/89, 2.5 ul will footprint
3. CT-2 F1 from 2/17/89, 2.5 ul will footprint
4. CT-2 F1 from 2/17/89, 2.5 ul will footprint
5. CT-2 F3 from 2/17/89, 2.5 ul footprints weakly
6. CT-2 F3 from 2/17/89, 2.5 ul footprints weakly
7. Mark Biggins GAGA factor from embryos, seems to have considerable nonspecific binding.
8. Mark Biggins GAGA factor from embryos, seems to have considerable nonspecific binding.
9. Recombinant GAGA from Howard Granouk, 2.5 ul footprints strongly.
10. Recombinant GAGA from Howard Granouk, 2.5 ul footprints strongly.
11. Recombinant GAGA from Howard Granouk, 2.5 ul footprints strongly.
12. GAGA fraction 4 from 6/18/91, 2.5 ul footprints strongly

DSG Proteins #1:
1. dTFIIB, recombinant, 300ug/ml from Wampler/Kadonaga
2. dTFIIE/F, natural from Wampler/Kadonaga
3. Gal4-VP16, from Workman's lab, Emanuel said 1:1000 dilution for footprints.
4. Gal4-VP16, from Workman's lab
5. 1:50 Gal4-VP16
6. 1:500 Gal4-VP16
7. 1:1000 Gal4-VP16

8-28. pol II, ~50ng/ul, Greenleaf-type prep, tube labelled at 6/28, see notebook 8/5/92, mostly IIb form.

29.

30. dTBP from Hoey/Tjian, 1992; 300 ng/ul, 30% pure
31. dTBP from Hoey/Tjian, 1992; 300 ng/ul, 30% pure
32. dTBP from Hoey/Tjian, 1992; 300 ng/ul, 30% pure
33. dTBP from Hoey/Tjian, 1992; 300 ng/ul, 30% pure
34.
35. dTBP from Emanuel, 9/9/92?, TBP 26
36. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
37. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
38. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
39. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
40. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
41. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
42. dTBP from Emanuel, 9/9/92?, TBP 26, probably 400 mM step
43.
44. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
45. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
46. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
47. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
48. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
49. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
50. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
51. dTBP from Emanuel, 9/9/92?, TBP 34, probably 1 M step
52.
53. dTBP from Emanuel, TBP16, 1 M step
54. dTBP from Emanuel, TBP16, 1 M step
55. dTBP from Emanuel, TBP16, 1 M step
56. dTBP from Emanuel, TBP17, 1 M step, adjacent fraction to previous one.

DSG Proteins #2:
This is a collection of P11 fractions from various sources.  Be careful to label the tubes when you remove them because it will be very easy to mix them up.  The collection was put together as people left the lab so there is no consistant labelling scheme.

1. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
2. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
3. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
4. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
5. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
6. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
7. P11 500, ~150 ul, P. Emanuel, used in G&D paper, Spring 94
8. 100 ul P11, 12/3/92, P. Emanuel, used in PNAS paper, Fall 93
9. 100 ul P11, 12/3/92, P. Emanuel, used in PNAS paper, Fall 93
10. TATA complex, (P11), 3/8/92, D. Gilmour, successfully used by many.
11. TATA complex, (P11), 3/8/92, D. Gilmour, successfully used by many.
12. TATA complex, (P11), 3/8/92, D. Gilmour, successfully used by many.
13. TATA complex, (P11), 3/8/92, D. Gilmour, successfully used by many.
14. TATA complex, (P11), 3/8/92, D. Gilmour, successfully used by many.
15. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
16. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
17. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
18. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
19. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
20. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94
21. TATA complex, (P11), 6/91, D. Gilmour, successfully used in gel shift 7/29/94

22-72. 4th P-11, TATA complex, M. Sypes, early 94, 2.8 mg/ml, 50% of the TAF250 appears to be degraded to 180 kd.  Successfully used in later part of Sypes thesis.

Bacteria Strains #1:
Notes: Glycerol Stocks, Stocks in slots 4 through 46 were prepared by Dev Idgunji

1. Mut L, competant cells from Washington University, for use in site directed mutagenesis.
2. hsp70 16mer clone in XBS, -194 to +84, from Mike Sypes.
3. *TATA hsp70 4mer clone, from Mike Sypes, probably in XBS.
4. JM109
5. JM109
6. HB101
7. HB101
8. HB101
9. HB101
10. HB101
11. HB101
12. HB101
13. HB101
14. HB101
15. HB101
16. HB101
17. HB101
18. HB101
19. HB101
20. DH5a
21. DH5a
22. DH5a
23. Car20ZT.2 -89
24. Car20ZT.2 -89
25. Car20ZT.2 -190
26. Car20ZT.2
27. Car20
28. Car20
29. pUC13
30. pUC13
31. P/T (hsp70 Promoter/Terminator)
32. P/T (hsp70 Promoter/Terminator)
33. P/E/T (hsp70 promoter/HA epitope/Terminator)
34. P/E/T (hsp70 promoter/HA epitope/Terminator)
35. PHET (hsp70 promoter/*C H1/HA epitope/Terminator)
36. PHET (hsp70 promoter/*C H1/HA epitope/Terminator)
37. Hsp70 S/X 16 WT from Beverly Purnell
38. Car20 -190 +89 (I'm not sure what they are because they don't have the ZT)
39. Car20 -190 +89 (I'm not sure what they are because they don't have the ZT)
40. Car20 -190 +89 (I'm not sure what they are because they don't have the ZT)
41. Car20 -190 +89 (I'm not sure what they are because they don't have the ZT)
42. Nuclear Localization Car20ZT.2 -89  (8/13/92; #1)
43. Nuclear Localization Car20ZT.2 -89  (8/13/92; #2)
44. Nuclear Localization Car20ZT.2 -89  (8/13/92; #3)
45. Nuclear Localization Car20ZT.2 (8/13/92; #4)
46. Nuclear Localization Car20ZT.2 (8/13/92; #5)
47.
48.
49.
50.
51.
52.
53.
54.
55.
56.
57.
58.
59.
60.
61.
62.
63.
64.
65.
66.
67.
68.
69.
70.
71.
72.
73. Gal4 1-94 (Joe Reese)
74. Gal4 1-147 VP16 (Joe Reese)
75. GPP78 (Chicca)
76. GPP93 (Chicca)
77. Gal4 1-147 (John Chicca)
78. Gal4 1-147 AH (Joe Reese)
79. Gal4-VP16 (John Chicca)
80.
81.

Autin's transcription factors 4/95 :

1. Dis79 S1 probe for Adh distal 10uM
2. pD-33CAT  Adh distal  0.68 ug/ul
3.
4. TBP 150ng/ul
5. TBP 150ng/ul
6. TBP 150ng/ul
7. TBP 150ng/ul
8. Pol II  2/10/95
9. Pol II  2/10/95
10. Pol II  2/10/95
11. Pol II  2/10/95
12. Pol II  2/10/95
13. Pol II  2/10/95
14. Pol II  2/10/95
15. IID (LS) 2/8/95
16. IID (LS) 2/8/95
17. IID (LS) 2/8/95
18. IID (LS) 2/8/95
19. IID (LS) 2/8/95
20. IID (LS) 2/8/95
21. IID (LS) 2/8/95
22. IIH 7/93
23. IIH 7/93
24. IIH 7/93
25. IIH 7/93
26. IIH 7/93
27. IIH 7/93
28. IIH 7/93
29. Prox71 S1 probe for Adh proximal, 18.6uM
30. pP*5'-55,  1.55 ug/ul
31. IIF 4/3/95
32. IIF 4/3/95
33. IIF 4/3/95
34. IIF 4/3/95
35. IIF 4/3/95
36.
37. rIIE 5/94
38.  rIIE 5/94
39.  rIIE 5/94
40.  rIIE 5/94
41.  rIIE 5/94
42.  rIIE 5/94
43. 5% PVA, 5%PEG
44.
45.
46.
47. rIIB 100ng/ul
48. rIIB 100ng/ul
49. rIIB 100ng/ul
 


Conaway's general factors:

Austin's 2nd shipment of factors: these have not been aliquoted.

Gilmour's TFIID.
This contains primarily tubes of Gilmour's DE52 fraction from 1997.  Chwen-Huey and Larry Benjamin have used these in binding studies.

Price/Greenleaf.
There are only a few tubes in this box.  Some KcN and DNase I inhibitor from David Price and the CTD-plus and CTD-minus versions of alph-amanitin resistant polymerase from Arno Greenleaf (this is the polymerase Baiyong used for his MCB paper).

RNA polymerase II.
This was prepared by Mike Sypes from NHS embryos.  The prep. is dated 6/17/99.  It was derived from 200 g of embryos.  An Elgin nuclear extract was passed over DEAE and the 0.6M fraction was collected.  This fraction was locaded on Heparin sepharose in 200 mM and eluted with 500 mM.  Next, the polII fraction was purified on MonoQ.  The final salt concentration is probably around 300 to 400 mM since it elutes as a sharp peak at around 500 mM.  The tubes have been labelled on top as "polII" in blue lettering.

Competent BL21(DE3) cells prepared by Jizhong Zou during his rotation in Fall 99.  They appear to be competent but the transformation efficiency is quite low.  Should be adequate for transforming with plasmids.  I suggests tranforming with 50 ng of plasmid.