AG Daniel:Freezer

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AG Daniel

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Freezer
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Forschungshaus Charité
Lindenberger Weg 80
13125 Berlin

Lab Members

General Freezer (-20 °C)

Name Cat. number Company Comment
Puromycin
Zeocin
M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant Promega
Bioline Bioscript Bioline
Bioline Random Hexamer Primers Bioline
Bioline RNase Inhibitors Bioline
Bioline 5x Reaction Buffer for Bioscript Bioline
Biotinylated Protein Ladder Detection Pack Cell Signaling
Proteinase K
Monoclonal Anti-Phosphoserine Clone BR-45 Sigma
InSolution ALLN
Alsterpaullone Calbiochem
Caspase Inhibitor 1 Calbiochem
Q-VD-oPh, Non-o-methylated Calbiochem
CdK1 Inhibitor CGP74514A Calbiochem
X-Gal Promega


Enzyme Freezer (-20 °C)

I-SceI Meganuclease kit

Klenow fragment

Klenow fragment


Polymerase

- GoTaq® Polymerase

GoTaq


- InviTaq Polymerase

InviTaq


- DreamTaq™ Green DNA Polymerase

    10x DreamTaq™ Green Buffer  5 µl
    dNTP Mix, 2mM each          5 µl (0.2 mM of each)
    Forward primer              0.1 - 1.0 µM
    Reverse primer              0.1 - 1.0 µM
    Template DNA                10 pg - 1 µg
    DreamTaq™ DNA Polymerase    1.25 u
    Water, nuclease-free        to 50µl
                  Total Volume  50µl


Shrimp Alkaline Phosphatase

Phosphatase catalyzes the removal of 5´ phosphate groups from DNA and RNA. Since phosphatase-treated fragments lack the 5´ phosphoryl termini required by ligases, they cannot self-ligate. This property can be used to decrease the vector background in cloning strategies.

USB SAP #E70092Y


T4 DNA Ligase

NEB T4 Ligase


TOPO® TA cloning kit

Invitrogen TOPO cloning


Restriction Enzymes

(status: 02/2010)

Name Restriction site NEBuffer 1 2 3 4 Comments
AccI 5'...GT MKAC...3'
3'...CAKM TG...5'
050 050 010 100
AfeI 5'...AGC GCT...3'
3'...TCG CGA...5'
025 050 025 100 blunt
AflII 5'...C TTAAG...3'
3'...GAATT C...5'
050 100 025 100 + BSA
AluI 5'...AG CT...3'
3'...TC GA...5'
100 100 075 100 blunt
ApaI 5'...GGGCC C...3'
3'...C CCGGG...5'
025 050 000 100 + BSA
ApaLI 5'...G TGCAC...3'
3'...CACGT G...5'
100 100 010 100 + BSA
AvaI 5'...C YCGRG...3'
3'...GRGCY C...5'
010 075 010 100
AvaII 5'...G GWCC...3'
3'...CCWG G...5'
050 075 010 100
AvrII 5'...C CTAGG...3'
3'...GGATC C...5'
100 100 050 100
BamHI 5'...G GATCC...3'
3'...CCTAG G...5'
075 100 100 100 + BSA
BbsI 5'...GAAGACNN NNNNNNNN...3' (N2)
3'...CTTCTGNNNNNN NNNN...5' (N6)
100 100 025 075
BclI 5'...T GATCA...3'
3'...ACTAG T...5'
050 100 100 075 [50°C]
BglII 5'...A GATCT...3'
3'...TCTAG A...5'
010 075 100 010
BspEI 5'...T CCGGA...3'
3'...AGGCC T...5'
000 010 100 000
BspHI 5'...T CATGA...3'
3'...AGTAC T...5'
010 100 050 100
BstEII 5'...G GTNACC...3'
3'...CCANTG G...5'
050 075 100 075 + BSA [60°C]
BstXI 5'...CCANNNNN NTGG...3'
3'...GGTN NNNNNACC...5'
025 100 100 050 [55°C]
Bsu36I 5'...CC TNAGG...3'
3'...GGANT CC...5'
000 025 100 000 + BSA
ClaI 5'...AT CGAT...3'
3'...TAGC TA ...5'
010 050 050 100 + BSA
EcoNI 5'...CCTNN NNNAGG...3'
3'...GGANNN NNTCC...5'
100 100 075 100
EcoRI 5'...G AATTC...3'
3'...CTTAA G...5'
100 100 100 100
EcoRV 5'...GAT ATC...3'
3'...CTA TAG...5'
100 100 100 100 + BSA blunt
HaeII 5'...RGCGC Y...3'
3'...Y CGCGR...5'
075 100 050 100 + BSA
HindIII 5'...A AGCTT...3'
3'...TTCGA A...5'
050 100 010 050
HinfI 5'...G ANTC...3'
3'...CTNA G...5'
075 100 075 075
HpaI 5'...GTT AAC...3'
3'...CAA TTG...5'
025 050 010 100 blunt
I-SceI 5'...TAGGGATAA CAGGGTAAT...3'
3'...ATCCC TATTGTCCCATTA...5'
010 050 050 050 + BSA
KpnI 5'...GGTAC C...3'
3'...C CATGG...5'
100 075 000 050 + BSA
MfeI 5'...C AATTG...3'
3'...GTTAA C...5'
075 050 010 100
MluI 5'...A CGCGT...3'
3'...TGCGC A...5'
025 075 100 050
NaeI 5'...GCC GGC...3'
3'...CGG CCG...5'
100 075 010 075 blunt
NcoI 5'...C CATGG...3'
3'...GGTAC C...5'
100 100 100 100
NdeI 5'...CA TATG...3'
3'...GTAT AC...5'
075 100 075 100
NheI 5'...G CTAGC...3'
3'...CGATC G...5'
100 100 010 100 + BSA
NotI 5'...GC GGCCGC...3'
3'...CGCCGG CG...5'
000 050 100 025 + BSA
NruI 5'...TCG CGA...3'
3'...AGC GCT...5'
000 010 100 010 blunt
NsiI 5'...ATGCA T...3'
3'...T ACGTA...5'
010 075 100 025
PacI 5'...TTAAT TAA...3'
3'...AAT TAATT...5'
100 075 010 100 + BSA
PmlI 5'...CAC GTG...3'
3'...GTG CAC...5'
100 075 000 075 blunt
PshAI 5'...GACNN NNGTC...3'
3'...CTGNN NNCAG...5'
050 075 010 100 + BSA blunt
PstI 5'...CTGCA G...3'
3'...G ACGTC...5'
075 075 100 050 + BSA
PvuI 5'...CGAT CG...3'
3'...GC TAGC...5'
010 075 100 010 + BSA
PvuII 5'...CAG CTG...3'
3'...GTC GAC...5'
100 100 100 100 blunt
SacI 5'...GAGCT C...3'
3'...C TCGAG...5'
100 050 010 100 + BSA
SacII 5'...CCGC GG...3'
3'...GG CGCC...5'
025 075 010 100
SalI 5'...G TCGAC...3'
3'...CAGCT G...5'
000 000 100 000 + BSA
SfiI 5'...GGCCNNNN NGGCC...3'
3'...CCGGN NNNNCCGG...5'
000 100 010 100 + BSA [50°C]
SmaI 5'...CCC GGG...3'
3'...GGG CCC...5'
000 000 000 100 [25°C] blunt
SnaBI 5'...TAC GTA...3'
3'...ATG CAT...5'
025 050 025 100 + BSA blunt
SpeI 5'...A CTAGT...3'
3'...TGATC A...5'
075 100 025 075 + BSA
SphI 5'...GCATG C...3'
3'...C GTACG...5'
100 100 050 100
SspI 5'...AAT ATT...3'
3'...TTA TAA...5'
050 100 050 050 blunt
StuI 5'...AGG CCT...3'
3'...TCC GGA...5'
100 100 050 100 blunt
SwaI 5'...ATTT AAAT...3'
3'...TAAA TTTA...5'
010 010 100 010 + BSA [25°C] blunt
Tth111 5'...GACN NNGTC...3'
3'...CTGNN NCAG...5'
100 025 025 100 [65°C]
XbaI 5'...T CTAGA...3'
3'...AGATC T...5'
000 100 075 075 + BSA DAM Methylierung!!!
XcmI 5'...CCANNNNN NNNNTGG...3'
3'...GGTNNNN NNNNNACC...5'
010 100 050 050
XhoI 5'...C TCGAG...3'
3'...GAGCT C...5'
075 100 100 100 + BSA
XmnI 5'...GAANN NNTTC...3'
3'...CTTNN NNAAG...5'
100 100 075 100 + BSA blunt


Name Restriction site NEBuffer 1 2 3 4 Comments
AatII 5'...GACGT C...3'
3'...C TGCAG...5'
000 050 050 100 Bernd
AscI 5'...GG CGCGCC...3'
3'...CCGCGC GG...5'
000 010 010 100 Bernd
DraI 5'...TTT AAA...3'
3'...AAA TTT...5'
075 075 050 100 Bernd
EagI 5'...C GGCCG...3'
3'...GCCGG C...5'
010 025 100 010 Bernd
MspI 5'...C CGG...3'
3'...GGC C...5'
075 100 050 050 Bernd
Psp1406I / AclI 5'...AA CGTT...3'
3'...TTGC AA...5'
010 010 000 100 + BSA Bernd
SbfI 5'...CCTGCA GG...3'
3'...GG ACGTCC...5'
075 050 000 100 Bernd
SfcI 5'...C TRYAG...3'
3'...GAYRT C...5'
075 050 010 100 + BSA Bernd
SrfI 5'...GCCC GGGC...3'
3'...CGGG CCCG...5'
 ??? Bernd
SrfII 5'...???...3'
3'...???...5'
 ??? Bernd


0402 Freezer (-20 °C)

Name Cat. number Company comment
Brefeldin A B-1009 A.G. Scientific
GeneJET PCR Purification Kit #K0701, #K0702 Fermentas
Zeocin Invitrogen
Salubrinal sc-202332 Santa Cruz
GoTaq Polymerase Promega


Name Restriction site NEBuffer 1 2 3 4 Comments
AflII 5'...C TTAAG...3'
3'...GAATT C...5'
050 100 025 100 + BSA
BamHI 5'...G GATCC...3'
3'...CCTAG G...5'
075 100 100 100 + BSA
EcoRI 5'...G AATTC...3'
3'...CTTAA G...5'
100 100 100 100
EcoRV 5'...GAT ATC...3'
3'...CTA TAG...5'
100 100 100 100 + BSA blunt
NdeI 5'...CA TATG...3'
3'...GTAT AC...5'
075 100 075 100
NheI 5'...G CTAGC...3'
3'...CGATC G...5'
100 100 010 100 + BSA
NotI 5'...GC GGCCGC...3'
3'...CGCCGG CG...5'
000 050 100 025 + BSA
NruI 5'...TCG CGA...3'
3'...AGC GCT...5'
000 010 100 010 blunt
PstI 5'...CTGCA G...3'
3'...G ACGTC...5'
075 075 100 050 + BSA
SacII 5'...CCGC GG...3'
3'...GG CGCC...5'
025 075 010 100
SfiI 5'...GGCCNNNN NGGCC...3'
3'...CCGGN NNNNCCGG...5'
000 100 010 100 + BSA [50°C]
SmaI 5'...CCC GGG...3'
3'...GGG CCC...5'
000 000 000 100 [25°C] blunt
StuI 5'...AGG CCT...3'
3'...TCC GGA...5'
100 100 050 100 blunt
XbaI 5'...T CTAGA...3'
3'...AGATC T...5'
000 100 075 075 + BSA DAM Methylierung!!!
XhoI 5'...C TCGAG...3'
3'...GAGCT C...5'
075 100 100 100 + BSA



S1 Freezer (-80 °C)

Rack no. Position 1 Position 2 Position 3 Position 4
01
02
03
04
05
Rack no. Position 1 Position 2 Position 3 Position 4
06 Tim Tim Tim Tim
07 Tim Tim Christian Herrberger Christian Herrberger
08 Christian Herrberger Christian Herrberger Tim Tim
09 Tim Tim Tim Tim
10 Tim Tim Tim Tim
Rack no. Position 1 Position 2 Position 3 Position 4
11
12
13
14
15
Rack no. Position 1 Position 2 Position 3 Position 4
16
17
18
19
20
Rack no. Position 1 Position 2 Position 3 Position 4
21
22
23
24
25
Rack no. Position 1 Position 2 Position 3 Position 4
26 ccDH5α, pUC19 DNA ccDH5α STOCK
27
28
29
30
Rack no. Position 1 Position 2 Position 3 Position 4
31
32
33
34
35
Rack no. Position 1 Position 2 Position 3 Position 4
36
37
38
39
40
Rack no. Position 1 Position 2 Position 3 Position 4
41
42
43
44
45


S1 Freezer #2 (-80 °C)

Rack no. Position 1 Position 2 Position 3 Position 4
01
02
03
04


S2 Freezer (-80 °C)

Status 01/2012

Rack no. Position 1 Position 2 Position 3 Position 4
01
02
03
04
Rack no. Position 1 Position 2 Position 3 Position 4
05
06
07
08
Rack no. Position 1 Position 2 Position 3 Position 4
09
10
11
12
Rack no. Position 1 Position 2 Position 3 Position 4
13
14
15 Jana; p73 Titer 2,5x105 Jana; ΔNp73 Titer 6,5x104
16 p53; Batch 52; Titer: 5x108
Rack no. Position 1 Position 2 Position 3 Position 4
17
18
19
20
Rack no. Position 1 Position 2 Position 3 Position 4
21
22
23 A&A; ARFtet Titer: 5x106 A&A; ΔNp14ARFtet Titer: 1.5x107
24
Rack no. Position 1 Position 2 Position 3 Position 4
25 p14ARF; #8; 05/2005 Annika & Philipp; LacZ; 06/05 p14ARF 1+2 p14ARF 1+2
26 CEANBK CEANBK #7.2 #7.7 RSVβGAL / CMVβGAL
27 Annika; p14ARF; 03.08.10; #25; Titer 2-3x107 Annika; p14ARF; 23.03.07; Titer 6x108 Annika; LacZ; 19.04.06; 1x108 Annika; PUMA; 01.05.07; Titer x10?
28 Annika; p14ARF Annika; p14ARF p14ARF; 7/7I; 7/7II; 7/7III p14ARF; 7/72; 04/08
Rack no. Position 1 Position 2 Position 3 Position 4
29 Tim; GFP; 11/08  ?; LacZ; 04/07 AHas; verschiedene BH3s Tim; CHOPtet; Titer: 8,2x105
30  ?; LacZ; T003; 03/06  ?; NBKtet; 01/2007 A&A; NBK A&A; NBK; 03/06
31  ?; verschiedenes  ?; ARF 03/07  ?; ARF 01/2007  ?; ΔNp14ARF; 03/06
32 Tim; LacZ; 11/08; Titer: 1x108  ?; Zellen EMPTY Tim ATF4tet; Titer 1,6x106


N2 tank (-196 °C)

Rack no. Position 1 Position 2 Position 3 Position 4
01
02
03
04
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