Team:TU-Munich/Notebook/Materials

From 2013.igem.org

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==Cell lines==
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*''E. coli'' XL-1 Blue (for molecular cloning)
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*''E. coli'' BL21 (for cytoplasmatic protein production
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*''P. patens'' clone 11 obtained from Prof. Reski (Freiburg University)
==Reagents==
==Reagents==
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<H3>Chemicals</H3>
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==Chemicals==
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{|
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|Reagent
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|Supplier
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! Reagent
 +
! Supplier
 +
! Location
 +
|-
 +
| 2-Mercaptoethanol
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
 +
| Acetonitrile (LC-MS grade)
 +
| Sigma-Aldrich
 +
| Taufkirchen, Germany
 +
|-
 +
| 30% Acrylamid with 0,8% Bisacrylamid
 +
| Serva Electrophoresis GmbH
 +
| Heidelberg, Germany
 +
|-
 +
| Agarose
 +
| peqlab Biotechnologie GmbH
 +
| Erlangen, Germany
 +
|-
 +
| Acetic acid (LC-MS grade)
 +
| Sigma-Aldrich Chemie GmbH
 +
| Taufkirchen
 +
|-
 +
| Ammonium acetate
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
 +
| Bacto Agar Becton
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| BD, DIFCO Laboratories GmbH
 +
| Augsburg, Germany
 +
|-
 +
| Bacto Tryptone Becton
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| BD, DIFCO Laboratories GmbH
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| Augsburg, Germany
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|-
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| Bacto Yeast Extract Becton
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| BD, DIFCO Laboratories GmbH
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| Augsburg, Germany
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|-
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|Biotin
 +
|
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|
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|-
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| Boric acid
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| AppliChem GmbH
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| Darmstadt, Germany
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|-
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| Bromphenol blue
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
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| Coomassie brilliant blue R-250
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
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|-
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| Chloramphenicol
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| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
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| D(+)-Glukose, monohydrate
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
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| dNTPs
 +
| Invitrogen AG
 +
| Basel, Switzerland
 +
|-
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| Ethanol p.p., 99,9%
 +
| Honeywell Deutschland AG
 +
| Neunkirchen, Germany
 +
|-
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| Glycerol
 +
| Carl Roth GmbH & Co. KG
 +
| Karlsruhe, Germany
 +
|-
 +
| Kaliumchloride (KCl)
 +
| Merck KG
 +
| Darmstadt, Germany
 +
|-
 +
| Kaliumdihydrogenphosphate (KH<sub>2</sub>PO<sub>4</sub>)
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
 +
| Methanol
 +
| Honeywell Deutschland AG
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| Neunkirchen, Germany
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|-
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| N,N,N’,N’-Tetramethylethylendiamin (TEMED)
 +
| AppliChem GmbH
 +
| Darmstadt, Germany
 +
|-
 +
| Sodiumchloride (NaCl)
 +
| Carl Roth GmbH & Co. KG
 +
| Karlsruhe, Germany
 +
|-
 +
| Sodiumdihydrogenphosphate (NaH2PO4)
 +
| Carl Roth GmbH & Co. KG
 +
| Karlsruhe, Germany
 +
|-
 +
| Disodiumhydrogenphosphate (Na2HPO4)
 +
| VWR International GmbH
 +
| Ismaning, Germany
 +
|-
 +
| Sodiumdodecylsulphate (SDS)
 +
| Carl Roth GmbH & Co. KG
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| Karlsruhe, Germany
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|-
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==References:==
 
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<div class="box-center">
 
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<!-- Kopiervorlagen -->
 
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[[http://www.ncbi.nlm.nih.gov/pubmed/6327079 Edens et al., 1984]]
 
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#[[http://www.ncbi.nlm.nih.gov/pubmed/6327079 Edens et al., 1984]] Edens, L., Bom, I., Ledeboer, A. M., Maat, J., Toonen, M. Y., Visser, C., and Verrips, C. T. (1984). Synthesis and processing of the plant protein thaumatin in yeast. ''Cell'', 37(2):629–33.
 
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<!-- Ab hier richtige Referenzen einfügen -->
 
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#
 
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Latest revision as of 01:47, 5 October 2013


Cell lines

  • E. coli XL-1 Blue (for molecular cloning)
  • E. coli BL21 (for cytoplasmatic protein production
  • P. patens clone 11 obtained from Prof. Reski (Freiburg University)

Reagents

Chemicals

Reagent Supplier Location
2-Mercaptoethanol AppliChem GmbH Darmstadt, Germany
Acetonitrile (LC-MS grade) Sigma-Aldrich Taufkirchen, Germany
30% Acrylamid with 0,8% Bisacrylamid Serva Electrophoresis GmbH Heidelberg, Germany
Agarose peqlab Biotechnologie GmbH Erlangen, Germany
Acetic acid (LC-MS grade) Sigma-Aldrich Chemie GmbH Taufkirchen
Ammonium acetate AppliChem GmbH Darmstadt, Germany
Bacto Agar Becton BD, DIFCO Laboratories GmbH Augsburg, Germany
Bacto Tryptone Becton BD, DIFCO Laboratories GmbH Augsburg, Germany
Bacto Yeast Extract Becton BD, DIFCO Laboratories GmbH Augsburg, Germany
Biotin
Boric acid AppliChem GmbH Darmstadt, Germany
Bromphenol blue AppliChem GmbH Darmstadt, Germany
Coomassie brilliant blue R-250 AppliChem GmbH Darmstadt, Germany
Chloramphenicol AppliChem GmbH Darmstadt, Germany
D(+)-Glukose, monohydrate AppliChem GmbH Darmstadt, Germany
dNTPs Invitrogen AG Basel, Switzerland
Ethanol p.p., 99,9% Honeywell Deutschland AG Neunkirchen, Germany
Glycerol Carl Roth GmbH & Co. KG Karlsruhe, Germany
Kaliumchloride (KCl) Merck KG Darmstadt, Germany
Kaliumdihydrogenphosphate (KH2PO4) AppliChem GmbH Darmstadt, Germany
Methanol Honeywell Deutschland AG Neunkirchen, Germany
N,N,N’,N’-Tetramethylethylendiamin (TEMED) AppliChem GmbH Darmstadt, Germany
Sodiumchloride (NaCl) Carl Roth GmbH & Co. KG Karlsruhe, Germany
Sodiumdihydrogenphosphate (NaH2PO4) Carl Roth GmbH & Co. KG Karlsruhe, Germany
Disodiumhydrogenphosphate (Na2HPO4) VWR International GmbH Ismaning, Germany
Sodiumdodecylsulphate (SDS) Carl Roth GmbH & Co. KG Karlsruhe, Germany

Kits and Standards

Plastic Disposables

Oligonucleotides

Name Sequence
EreA_for 5’- GCGCGTCTAGATGGCCGGCACGTGGAGAACGACCAGA -3‘
EreA-rev 5‘- TTCGAAACCGGTTAGGGCAACTAGGCTGTCC -3‘
EreA_SDMI_for 5’- CGAATCAGGAAGAAAACTACAGTTAGTCGGAATCGAC -3‘
EreA_SDMI_rev 5'- GTCGATTCCGACTAACTGTAGTTTTCTTCCTGATTCG -3'
EreA_SDMII_for 5'- GACGCCTGTGGTACGGAAAATTCATGTCTGACATT - 3'
EreA_SDMII_rev 5'- AATGTCAGACATGAATTTTCCGTACCACAGGCGTC - 3'
EreA_SDMIII_for 5'- GTGTTCCAAGCGCGGGCAAGGACAGCC - 3'
EreA_SDMIII_rev 5'- GGCTGTCCTTGCCCGCGCTTGGAACAC - 3'
EreB_for 5'- GCGCGTCTAGATGGCCGGCAGGTTCGAAGAATGGGTC - 3'
EreB-rev 5'- TTCGAAACCGGTTTCATAAACGACCTCAGATACAG - 3'
EreB_SDMI_for 5'- GGTGAAAATTCTCATTTCATAAAAGAGTTCTTTTTGTTACGACATACGCTTTT - 3'
EreB_SDMI_rev 5'- AAAAGCGTATGTCGTAACAAAAAGAACTCTTTTATGAAATGAGAATTTTCACC - 3'
EreB_SDMII_for 5'- CTTATAGTGGGCATACAGCAGCCCTCTATCCGG - 3'
EreB_SDMII_rev 5'- CCGGATAGAGGGCTGCTGTATGCCCACTATAAG - 3'
EreB_SDMIII_for 5'- CGAGTTGATAACTTCCAACTACAGGAACCAAATGAAGGTTC - 3'
EreB_SDMIII_rev 5'- GAACCTTCATTTGGTTCCTGTAGTTGGAAGTTATCAACTCG - 3'
TEV_fw 5'- GCGCGTCTAGATGGCCGGCGGAGAAAGCTTGTTTAAGGGGC - 3'
TEV_rv 5'- ATATAACCGGTATTCATGAGTTGAGTCGCTTCC - 3'
TEV_t387c_fw 5'- ATGGTGTCAGACACTAGCTGCACATTCCCTTCATC - 3'
TEV_t387c_rv 5'- GATGAAGGGAATGTGCAGCTAGTGTCTGACACCAT - 3'
N_split_TEV_fw 5'- ATTCGCGGCCGCTTCTAG - 3'
N_split_TEV_rv 5'- TTCGAAACCGGTAGTTTGGAAGTTGGTTGTCAC - 3'
C_split_TEV_fw 5'- AAGCTTTCTAGATGGCCGGCAAGAGCATGTCTAGCATGGTG - 3'
C_split_TEV_rv 5'- CCGCTACTAGTATTAACCGG - 3'
PIF6_2-100_fw 5'- AAGCTTTCTAGATGGCCGGCATGTTCTTACCAACCGATTATTGTTGC - 3'
PIF6_2-100_rv 5'- TTCGAAACCGGTGTCAACATGTTTATTGCTTTC - 3'
MinMCS_fo 5'- CTAGACCTAGGAAGCTTCCTGCAGGACTAGTAAGCTTCTGCA - 3'
MinMCS_rev 5'- GAAGCTTACTAGTCCTGCAGGAAGCTTCCTAGGT - 3'
Bpu_for 5'- GCGCGTCTAGATGGCCGGCAACCTAGAAAAATTTGTTGACGAGCTG - 3'
Bpu_rev 5'- TTCGAAACCGGTGATGATATCCATCGGCCGCAT - 3'
Bpu_SDMI_for 5'- CACTTAAGCATTTTCTACGGGTCGATCACACCATTCA - 3'
Bpu_SDMI_rev 5'- TGAATGGTGTGATCGACCCGTAGAAAATGCTTAAGTG - 3'
Bpu_SDMII_fo 5'- GACACGATTAAATGTGTATGCGGGCTTAGCTGGATTTTATTTG - 3'
Bpu_SDMII_re 5'- CAAATAAAATCCAGCTAAGCCCGCATACACATTTAATCGTGTC - 3'
Bpu_SDMIII_fo 5'- CACCCTTAAAAATAAAGCTGGCTGCGGACAGGAAG - 3'
Bpu_SDMIII_re 5'- TTCCTGTCCGCAGCCAGCTTTATTTTTAAGGGTG - 3'
Bpu_SDMIV_fo 5'- CGTACGCTCACTCTTACGGGTACACAGGATAAATA - 3'
Bpu_SDMIV_re 5'- TATTTATCCTGTGTACCCGTAAGAGTGAGCGTACG - 3'
xylE_for 5'- AAGCTTTCTAGATGGCCGGCAACAAAGGTGTAATGCGACCG - 3'
xylE_rev 5'- TTCGAAACCGGTGGTCAGCACGGTCATGAATC - 3'
xylE_c312a_for 5'- GAACAGTTGTGGACGGCGCGTGCGC - 3'
xylE_c312a_rev 5'- GCGCACGCGCCGTCCACAACTGTTC - 3'
xylE_g483a_for 5'- CCTCATGTATGGCGACGAATTACCGGCGACCTA - 3'
xylE_g483a_rev 5'- TAGGTCGCCGGTAATTCGTCGCCATACATGAGG - 3'
xylE_a834g_for 5'- CAACTACCCGGACCACAAGCCGGTGACCT - 3'
xylE_a834_rev 5'- AGGTCACCGGCTTGTGGTCCGGGTAGTTG - 3'
p Act_for 5'- AAGCTTTCTAGAGTCGACGAGGTCATTCATATG - 3'
pAct_rev 5'- CTCGAGCTGCAGCGGCCGCTACTAGTAGCTTCTACCTACAAAAAAGCTC - 3'
t35S_for 5'- AAGCTTTCTAGAGCTAGAGTCCGCAAAAATCACC - 3'
t35S_rev 5'- CTCGAGCTGCAGCGGCCGCTACTAGTAGGTCACTGGATTTTGGTTTTAGG - 3'
Npt_for 5'- AAGCTTGAATTCGCGGCCGCTTCTAGAGATCGGATCCTGTCAAACAC - 3'
Npt_rev 5'- CGGCCGCTACTAGTAATCAAGCTTGACAGGAGGC - 3'
miniMCSII_fw 5'- CTAGACAATTGAAGCTTCCTGCAGGACTAGTAAGCTTCTGCA - 3'
miniMCSII_rv 5'- GAAGCTTACTAGTCCTGCAGGAAGCTTCAATTGT - 3'
SpyTag_fw 5'- CTAGATGGCCGGCGCTCATATTGTCATGGTTGATGCTTACAAGCCAACTAAGA - 3'
SpyTag_rv 5'- CCGGTCTTAGTTGGCTTGTAAGCATCAACCATGACAATATGAGCGCCGGCCAT - 3'
PIF3(2-100)_fw 5'- AAGCTTTCTAGATGGCCGGCCCTCTGTTTGAACTTTTCAGG - 3'
PIF3(2-100)_rv 5'- CCGCTACTAGTATTAACCGG - 3'
NucA_fw 5'- AAGCTTTCTAGATGGCCGGCGCAACTTCAACTAAAAAATTACATAAAGAACCTG - 3'
NucA_rv 5'- TTCGAAACCGGTTTGACCTGAATCAGCGTTGTC - 3'
AlcR_fw 5'- AAGCTTGAATTCGCGGCCGCTTCTAGATGGCAGATACGCGCCGAC - 3'
AlcR_rv 5'- TTCGAAACTAGTACTACAAAAAGCTGTCAACTTTCCCATTCAAACC - 3'
SQ_pAutoRex8_1 5'- CGGGGCATCACAGCGTCG - 3'
SQ_pAutoRex8_2 5'- TATCGGTAACCGAGTCCG - 3'
AlcR_QCI_fw 5'- GGCTAAATGTTCCTGCTGACATAGCCGCCGATG - 3'
AlcR_QCI_rv 5'- CATCGGCGGCTATGTCAGCAGGAACATTTAGCC - 3'
AlcR_QCII_fw 5'- GCACGGTATGGGTGCACGTTTGGACAAGCTACTCGAAAATG - 3'
AlcR_QCII_rv 5'- CATTTTCGAGTAGCTTGTCCAAACGTGCACCCATACCGTGC - 3'
AlcR_QCIII_fw 5'- CATCCAGTCTTGGTACGTCATTTTGGACGGTCACTGGCAT - 3'
AlcR_QCIII_rv 5'- ATGCCAGTGACCGTCCAAAATGACGTACCAAGACTGGATG - 3'
AlcR_QCIV_fw 5'- GCCGCAGCTGCAAGTACCCCGCTAG - 3'
AlcR_QCIV_rv 5'- ATTTTTGAGAAAAATATATATTCAGGCGAATTGCACAATGAACAATAATAAGATTAAAATAG - 3'
pActin_QC_fw 5'- ATTTTTGAGAAAAATATATATTCAGGCGAATTGCACAATGAACAATAATAAGATTAAAATAG - 3'
pActin_QC_rv 5'- CTATTTTAATCTTATTATTGTTCATTGTGCAATTCGCCTGAATATATATTTTTCTCAAAAAT - 3'
NPT_QCI_fw 5'- GTCCGGTGCCCTGAATGAATTGCAGGACGAG - 3'
NPT_QCI_rv 5'- CTCGTCCTGCAATTCATTCAGGGCACCGGAC - 3'
NPT_QCII_fw 5'- GCGGGACCCAAGCTTTAGATCTTGCTGCG - 3'
NPT_QCII_rv 5'- CGCAGCAAGATCTAAAGCTTGGGTCCCGC - 3'
IRES_polio_fw 5'- AAGCTTGAATTCGCGGCCGCTTCTAGAGTTAAAACAGCTCTGGGGTTG - 3'
IRES_polio_rv 5'- TTGCAACTGCAGCGGCCGCTACTAGTAAATCCAATTCGCTTTATGATAACAATC - 3'
BPU_SDMI_r_for 5'- CCACTTAAGCATTTTCTACCAGTCGATCACACCATTCACG - 3'
BPU_SDMI_r_rev 5'- CGTGAATGGTGTGATCGACTGGTAGAAAATGCTTAAGTGG - 3'
Strep-TEV_for 5'- CTAGATGGCCGGCAGCGCTTGGTCTCATCCTCAATTTGAAAAGGGTGAAAATCTTTATTTTCAAAGCGGAA - 3'
Strep-TEV_rev 5'- CCGGTTCCGCTTTGAAAATAAAGATTTTCACCCTTTTCAAATTGAGGATGAGACCAAGCGCTGCCGGCCAT - 3'
SQ_pluc-Actin_1 5'- CCGACTCAAATACAGATATGC - 3'
SQ_pluc-Actin_2 5'- CTCGGATGTAGATCTGCG - 3'
pActin_QC_ins_fw 5'- ACCCCCCCCTCTCCTCCCATCCCCCCAACCCTACCACCACCACC
ACCACCACCTCCACCTCCTCCCCCCTCGCTGCCGGACG - 3'

pActin_QC_ins_rv 5'- CGTCCGGCAGCGAGGGGGGAGGAGGTGGAGGTGGTGGTGGTGGT
GGTGGTAGGGTTGGGGGGATGGGAGGAGAGGGGGGGGT - 3'

pActin_Oryza_fw 5'- AAGCTTTCTAGAGTAGCTAGCATACTCGAGGTCATTCATATGCTTG - 3'
pActin_Oryza_rv 5'- TTCGAACTGCAGCGGCCGCTACTAGTACTTCTACCTACAAAAAAGCTCCG - 3'
GGGGSx5_TEV_fw 5'- CTAGATGGCCGGCGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGGAGG
CGGTGGCTCCGGAGGTGGAGGCTCAGGAGAGAATTTGTATTTTCAGTCAGGAA - 3'

GGGGSx5_TEV_rv 5'- CCGGTTCCTGACTGAAAATACAAATTCTCTCCTGAGCCTCCACCTCCGGAGCCACCGCCTCCC
GATCCGCCACCGCCAGAGCCACCTCCGCCTGAACCGCCTCCACCGCCGGCCAT - 3'

mMCSII_fw 5'- AATTCGCGGCCGCTTCTAGAGCAATTGAAGCTTCCTGCAGGTA - 3'
mMCSII_rv 5'- CTAGTACCTGCAGGAAGCTTCAATTGCTCTAGAAGCGGCCGCG - 3'
FluA_tripmut_fw 5'- AAGCTTTCTAGATGGCCGGCGACGTGTACCACGACGGTG - 3'
FluA_tripmut_rv 5'- TTCGAAACCGGTATTGTTGACCTTGCAGGCGG - 3'
pActin_QCI_fw 5'- CATAATTTTTGAGAAAAATATATATTCAGGCGAATTGTCACAATGAACAATAATAAGATTAAAATA - 3'
pActin_QCI_rv 5'- TATTTTAATCTTATTATTGTTCATTGTGACAATTCGCCTGAATATATATTTTTCTCAAAAATTATG - 3'
Alc R fw Seq. 5'- CAGAGCACAATTGTCCATAC - 3'
Alc R rv Seq. 5'- TCGCGGTCGATGCTCTCC - 3'
pActin_QCins_fw 5'- CATCGCCACTATATACATACCCCCCCCTCTCCTCCCATCCCCCCAACCCTACCACCACCACCAC
CACCACCTCCACCTCCTCCCCCCTCGCTGCCGGACGACGAGCTCCTC - 3'

pActin_QCins_rv 5'- GAGGAGCTCGTCGTCCGGCAGCGAGGGGGGAGGAGGTGGAGGTGGTGGTGGTGGTGGTGGTAGGG
TTGGGGGGATGGGAGGAGAGGGGGGGGTATGTATATAGTGGCGATG - 3'

PP1_for 5'- GCCGCTTCTAGATGGCCGGCGGATCCGCGGATTTAGATA - 3'
PP1_rev 5'- GTATTAACCGGTTTTCTTTGCTTGCTTTGTGATC - 3'
PhyB fw Seq 5'- GCGATTTCTCAGTTACAGGC - 3'
PhyB rv Seq. 5'- CTCAAAGCACGAGAAAGAAGC - 3'
pAct_ins_fw 5'- ACCACCACCACCTCCACCTCCTCCCCCCTCGCTGCCGGACGACGAGCTC - 3'
pAct_ins_rv 5'- GGTGGTGGTAGGGTTGGGGGGATGGGAGGAGAGGGGGGGGTATGTATATAGTGGCGATGG - 3'
PppAct_fw 5'- AAGCTTGAATTCGCGGCCGCTTCTAGATCGAGTCCTTGTGACTTTTGTGCAT - 3'
PppAct_rv 5'- TTCGAAACTAGTATATTTAATACGGACCTGCACAACAACCACCAAT - 3'
PppActin_QC1_fw 5'- TTATTGAGGATATCCATCTATCTTGATTTTGGACAATGTTTTACTGC - 3'
PppActin_QC1_rv 5'- GCAGTAAAACATTGTCCAAAATCAAGATAGATGGATATCCTCAATAA - 3'
PppActin_QC2_fw 5'- ACACTCTACATTCATGTCAAGAGTATAGGGACTTGGG - 3'
PppActin_QC2_rv 5'- CCCAAGTCCCTATACTCTTGACATGAATGTAGAGTGT - 3'
PppActin_fw_corr 5'- AAGCTTGAATTCGCGGCCGCTTCTAGAGCGAGTCCTTGTGACTTTTGTGCAT - 3'
SYFP2_fw 5'- AAGCTTTCTAGATGGCCGGCGTTAGCAAGGGCGAAGAAC - 3'
SYFP2_rv 5'- TTCGAAACCGGTTTTATACAGCTCATCCATACCCAG - 3'
eCFP_fw 5'- AAGCTTTCTAGATGGCCGGCGTGAGCAAGGGCGAGGAG - 3'
eCFP_rv 5'- TTCGAAACCGGTCTTGTACAGCTCGTCCATGCC - 3'
G-TEV-site-G_fw 5'- CTAGATGGCCGGCGGAGAGAATTTGTATTTTCAGTCAGGAA - 3'
G-TEV-site-G_fw 5'- CCGGTTCCTGACTGAAAATACAAATTCTCTCCGCCGGCCAT - 3'
PppActin_seq_fw 5'- CATGTCCCAGGGCCCTAG - 3'

Instruments

Electrophoresis

Photometers

Nanodrop Spectrohpotometer

Moss-cultivation