RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1236

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_0524600; Gene model (P.falciparum): Not available; Gene product: BIR protein; PIR protein (PIR8) Name tag: mCherry
Phenotype	Asexual bloodstage; Liver stage;

Last modified: 18 November 2016, 13:05

*RMgm-1236

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Reference 1 (PMID number) : 27851824
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	P. berghei ANKA cl15cy1
Other information parent line	A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
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The mutant parasite was generated by
Name PI/Researcher	A. Fougère, C.J. Janse, B.M.D. Franke-Fayard
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center
City	Leiden
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-1236
Principal name	2312, 2313
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	The tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not different from wild type
Sporozoite	Not different from wild type
Liver stage	Expressed in liver stages (location in the cytoplasm of the parasite)
Additional remarks phenotype	Mutant/mutation Protein (function) Phenotype Additional information The mutant parasite expresses an mCherry-tagged protein.The protein has been selected for tagging in a screen for putative exported proteins of P. berghei. The genotype of the parasites has not been analysed in detail. Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) has been performed (see below). This analysis provided evidence for tagging of the correct gene. The construct used aims at integration of the tagging construct by single-cross-over integration resulting in the presence of a tagged copy of the endogenous gene. Phenotype analyses of the blood stages by fluorescence microscopy show that the tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm. Phenotype analyses of mosquito and liver stages showed no expression in mosquito stages (oocysts; sporozoites) whereas the protein was detected in the cytoplasm of maturing liver stages. Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_0524600

Gene Model P. falciparum ortholog Not available

Gene product BIR protein; PIR protein

Gene product: Alternative name PIR8

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Details of the genetic modification

Name of the tag mCherry

Details of tagging C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct (Linear) plasmid single cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

CATTCAAGACAAAATTCAATCTCTATCAATAAACTTTGTGTACTTTTGGGAATGTTCAAA

AAAAATGTCATTGTGTGAATTACCACCAGAAGAATTGAGACAAATATTCCAGAAATATTT

AGATGAACTATATCGTCTATTTTATTATTATATAAATATATCATTAATTTATGCAATATC

CCATGCAAGTGATGAAAAGGATGTAAAATATTATATACCATATTATGAAAAATTTTCACA

AAAATATAATGAAATAGCACAAAAATGTAATCTTTATAAATGCAGCCCATATTGTAAAGA

ATTATATAATTTGGGAAACGAATATTATAATGCTCGGAATGAATTTCTTCTACGTAATAG

AATATATATAAATAAATTACCAACATTATTCAAAATACCACCATGTTTTAAATCCAATAA

TAATAATGAACATTTGGAATGTGAAAATACGAAATATTTGAAACAGGAAAATAATTCAAA

TGATAAAATATTTAACTCACGAGTTTTATTTTCATTTATATATATACTTATTCCATTTTT

CTTATTAATAATTTATAAGGTAAAAACAATTATACTTCAAAAAAATTTTATTTTCATTTG

TTTTTTTTTGTGTTTACTATTGTTTAAATTTATCGGTTCCTTATATATTTATATATTTTT

TTTTCATAATAACAGTTTACATCCTTTGAGCTTTGGATGAATTCTATATTTAGAACTGAG

ATCGCACTTTCAGACAAAATAGAAGAAAATATTCAAGAGGAAAAAAATATTATGGAAAAA

GGCACCGATGAAGGAAATAAGGAAAATCAAGTGACAGAAAAAAATGGAAAAGATAAAAAA

AATGAAGAGCATCCAGAGTCAAATAATAACAAAGGATCCGGTGGAGGTGGAAGATCCATG

GTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTG

CACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGC

CCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTC

GCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCC

GCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTG

ATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGC

GAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATG

CAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCC

CTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAG

GTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAAC

ATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGC

GCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATT

AATATATATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTA

ATTTTTATATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTG

AAAAAATTCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCAT

AAAGTACGAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAAT

TTTATTCTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTAT

TGAAAATAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAA

AGTAAAGGGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAG

GAAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAAC

TATGCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAA

GAGACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTT

TCAAACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGAT

TTAATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTT

TTCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAA

AAAAAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAA

ATATATAAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATC

TCCATCTGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAA

TCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGT

TCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCC

ATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAA

AGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATAC

ATGATGCACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGC

GGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCT

GTTGGATTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTG

TAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCC

GGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTC

ACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCG

CGAATGAACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGA

AGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCA

ACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGG

TACTGGAACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTT

TGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCG

GCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAG

CTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGA

ACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGA

TACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGAC

TCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGG

AACGCAGGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGA

GAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATC

TGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCA

CAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCT

TTGAGGGAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGC

ATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGA

GAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGA

CTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAG

TGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACC

AGACACACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAA

GCAATATACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATA

GTCAATTTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTG

TATATAAAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTT

ACTATTTATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATT

TAATATTATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACT

TTTTTATTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATT

AATTCGCTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCT

CTTTTTCAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTAT

ATCCTACCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAAT

ATATAAATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTT

AAAATTGATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATA

ATACAGTGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTT

AATTGCCTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGC

TCCATTTTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATAT

ACTCATAAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCA

TTTTGTATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTT

GTACATGTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTT

TTAAATTTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTAT

ATATTTTTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAAC

TCTCTTTGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATA

TATATTATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTA

TTGGCATAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAAC

TATATCCGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATA

TTAAATAAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAA

GAGACCATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATT

GAGCATATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATA

AATTATTGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATA

TAGTTCATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTAT

ATATATATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAG

GAAAAAATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAA

ATATGTTAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAA

CAAAATGACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCC

AAAGAAAAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATAT

ATATTTATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCA

TACACAAACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGT

TATTATTATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATT

AAAAATAATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTG

CAGGCATGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTG

TTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGT

GTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAA

AGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGC

TTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAG

AGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGT

CGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGA

ATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCG

TAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAA

AAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTT

TCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCT

GTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCT

CAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCC

CGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTT

ATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGC

TACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTAT

CTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAA

ACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAA

AAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGA

AAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCT

TTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGA

CAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATC

CATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGG

CCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAAT

AAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCAT

CCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCG

CAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTC

ATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAA

AGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATC

ACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTT

TTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAG

TTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGT

GCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAG

ATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCAC

CAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGC

GACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCA

GGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGG

GGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTAAATTGTAAGCGTTAATATTTTGTTA

AAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGC

AAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGG

AACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTAT

CAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGC

CGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAG

CCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTG

GCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTA

CAGGGCGCGTCCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGG

GCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGG

GTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAGCGCGCGTAA

TACGACTCACTATAGGGCGAATTGGAGCTCCACCGCGGTGGCGGCCGCTCTAGAACTAGT

Restriction sites to linearize plasmid SnaBI

Selectable marker used to select the mutant parasite tgdhfr

Promoter of the selectable marker pbdhfr

Selection (positive) procedure pyrimethamine

Selection (negative) procedure No

Additional remarks genetic modification

Additional remarks selection procedure

Primer information: Primers used for amplification of the target sequences Click to view information

Primer information: Primers used for amplification of the target sequences Click to hide information

Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

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