Affinity Purification:Article Title: Chromatin-associated cohesin turns over extensively and forms new cohesive linkages in Drosophila oocytes during meiotic prophase.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies High-affinity rat monoclonal anti-HA (clone 3F10) Roche Cat#11863423001 Lot no. 14223800; RRID: AB_390918 Mouse monoclonal anti-C3G (clone 1A8-1G2) Hawley Lab47 N/A Rabbit anti-CID Rogers Lab48 N/A GFP-Booster Alexa Fluor 488 (Alpaca anti-GFP) ChromTek Cat#gb2AF488; RRID: AB_2827573 RFP-Booster Alexa Fluor 568 (Alpaca anti-mCherry) ChromTek Cat#rb2AF568; RRID: AB_2827576 Affinity-purified guinea pig anti-Smc1 Bickel Lab26 N/A Rabbit anti-GFP Invitrogen Cat#A11122; RRID: AB_221569 Monoclonal mouse anti-alpha-tubulin (12G10) Developmental Studies Hybridoma Bank (deposited by J. Frankel and E.M. Nelson) Cat#AB_1157911; RRID:N/A Cy3 anti-rat (min-X mouse) Jackson ImmunoResearch Labs Cat#712-165-153; RRID: AB_2340667 Cy5 anti-rabbit Jackson ImmunoResearch Labs Cat#711-175-152; RRID: AB_2340607 Alexa 488 anti-mouse (min-X rat) Jackson ImmunoResearch Labs Cat#715-545-151; RRID: AB_2341099 Alkaline phosphatase goat anti-rat Sigma-Aldrich Cat#A8438; RRID: AB_258391 Alkaline phosphatase goat anti-mouse Invitrogen Cat#A16069; RRID: AB_2534742 Alkaline phosphatase goat anti-guinea pig Southern Biotech Cat#6090-04; RRID: AB_2796156 Alkaline phosphatase goat anti-rabbit Promega Cat#S373B; RRID: AB_430872 Chemicals, peptides, and recombinant proteins Formaldehyde Ted Pella Cat#18505 NP-40 Thermo Fisher Cat#28324 Tween-20 (for cytology) Thermo Fisher Cat#28320 BSA Fisher Scientific Cat#BP1605-100 DAPI Invitrogen Cat#D1306 Poly-L-lysine Sigma-Aldrich Cat#P8920 SlowFade Diamond Antifade Molecular Probes Cat#S36967 Prolong Gold Antifade Molecular Probes Cat#P36930 Vectashield Vibrance Vector Laboratories CatH-1700 RIPA buffer Sigma-Aldrich Cat#R0278 HALT protease inhibitor Thermo Fisher Cat#87786 Benzonase Millipore Cat#E1014 EDTA Thermo Fisher Cat#1861274 Tween-20 (for immunoblots) BioRad Cat#1610781 Centrifugal filter Millipore Cat#UFC30GV00 7.5% protein gel BioRad Cat#4568023 Precision Plus Protein Dual Color Standards BioRad Cat#1610374 Immobilon-P membrane Millipore Cat#IPVH00010 AP immune star substrate BioRad Cat#1705018 Experimental models: Organisms/strains Drosophila stocks used in this study, see Table S1 This study N/A Oligonucleotides Alexa 647-labeled Oligopaint probe (OPP122), Mixture of 80-base oligos targeting 100kb distal region of the X chromosome (dm6, nucleotides 1,400,000-1,500,000) Joyce Lab, University of Pennsylvania N/A (Continued on next page) Current Biology 34, 1–12.e1–e6, July 8, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cy3-conjugated probe (50-Cy3-AGGGATCGTTAGCACTCGTAAT) hybridizes to 359-bp repeat in pericentric heterochromatin of the X chromosome Integrated DNA Technologies N/A Smc3* Fwd: 5’GACAAGACGCGCCGCACGCTAG AATACATTCGGTATGAGACCGAGCTGAAGGACA Integrated DNA Technologies N/A Smc3* Rev: 5’TGTCCTTCAGCTCGGTCTCATACCG AATGTATTCTAGCGTGCGGCGCGTCTTGTC Integrated DNA Technologies N/A HA stop Fwd: 5’GCTTCTAGCGAAAAAGTGTA GACGGTATCGATAAGCTTG Integrated DNA Technologies N/A HA stop Rev: 5’CAAGCTTATCGATACCGTC TACACTTTTTCGCTAGAAGC Integrated DNA Technologies N/A Smc3-WT Fwd: 5’AGCACTGGTACCGCGGCCG CAATAAGATGCACATCAAGCAG Integrated DNA Technologies N/A Smc3-WT Rev: 5’ACTGATCTCGAGGCGGCG TGGGTGCTGTCG Integrated DNA Technologies N/A Smc3-KK Fwd: 5’CGCGTGATTGGCGCCAGAA GGGACCAGTACTTCC Integrated DNA Technologies N/A Smc3-KK Rev: 5’GGAAGTACTGGTCCCTTCTGG CGCCAATCACGCG Integrated DNA Technologies N/A Recombinant DNA Clone RE14758 (Smc3 cDNA) DGRC Stock 8520; RRID:DGRC_8502 pUASP-attB Gift from Hawley lab49 RRID: N/A w+ attB vector Addgene (donated by Jeff Sekelsky) Plasmid# 30326; RRID: Addgene_30326 Software and algorithms Volocity Visualization, Restoration, and Quantitation Quorum Technologies, Canada Version 6.5.0 https://www.volocity4d.com/ downloadhttps://www. volocity4d.com/download Volocity Acquisition (for epifluorescence imaging) Quorum Technologies, Canada Version 6.5.1 https://www.volocity4d.com/ volocity-acquisition Nikon Elements (for spinning disc confocal imaging) Nikon Version 5.11.02 Build 1369 MATLAB Mathworks Version R2022a https://www.mathworks.com/ products/new_products/ release2022a.html BioRad Image Lab BioRad Version 6.1 https://www.bio-rad.com/ en-us/product/image-labsoftware?ID=KRE6P5E8Z Microsoft Office Microsoft Version 16.84 Affinity Designer Affinity Version 1.10.6.1665 https://affinity.serif.com/ en-us/designer/ Other Chemi Doc Touch Imaging system BioRad Serial no. 732BR0120 e2 Current Biology 34, 1–12.e1–e6, July 8, 2024 Article
Recombinant:Article Title: Chromatin-associated cohesin turns over extensively and forms new cohesive linkages in Drosophila oocytes during meiotic prophase.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies High-affinity rat monoclonal anti-HA (clone 3F10) Roche Cat#11863423001 Lot no. 14223800; RRID: AB_390918 Mouse monoclonal anti-C3G (clone 1A8-1G2) Hawley Lab47 N/A Rabbit anti-CID Rogers Lab48 N/A GFP-Booster Alexa Fluor 488 (Alpaca anti-GFP) ChromTek Cat#gb2AF488; RRID: AB_2827573 RFP-Booster Alexa Fluor 568 (Alpaca anti-mCherry) ChromTek Cat#rb2AF568; RRID: AB_2827576 Affinity-purified guinea pig anti-Smc1 Bickel Lab26 N/A Rabbit anti-GFP Invitrogen Cat#A11122; RRID: AB_221569 Monoclonal mouse anti-alpha-tubulin (12G10) Developmental Studies Hybridoma Bank (deposited by J. Frankel and E.M. Nelson) Cat#AB_1157911; RRID:N/A Cy3 anti-rat (min-X mouse) Jackson ImmunoResearch Labs Cat#712-165-153; RRID: AB_2340667 Cy5 anti-rabbit Jackson ImmunoResearch Labs Cat#711-175-152; RRID: AB_2340607 Alexa 488 anti-mouse (min-X rat) Jackson ImmunoResearch Labs Cat#715-545-151; RRID: AB_2341099 Alkaline phosphatase goat anti-rat Sigma-Aldrich Cat#A8438; RRID: AB_258391 Alkaline phosphatase goat anti-mouse Invitrogen Cat#A16069; RRID: AB_2534742 Alkaline phosphatase goat anti-guinea pig Southern Biotech Cat#6090-04; RRID: AB_2796156 Alkaline phosphatase goat anti-rabbit Promega Cat#S373B; RRID: AB_430872 Chemicals, peptides, and recombinant proteins Formaldehyde Ted Pella Cat#18505 NP-40 Thermo Fisher Cat#28324 Tween-20 (for cytology) Thermo Fisher Cat#28320 BSA Fisher Scientific Cat#BP1605-100 DAPI Invitrogen Cat#D1306 Poly-L-lysine Sigma-Aldrich Cat#P8920 SlowFade Diamond Antifade Molecular Probes Cat#S36967 Prolong Gold Antifade Molecular Probes Cat#P36930 Vectashield Vibrance Vector Laboratories CatH-1700 RIPA buffer Sigma-Aldrich Cat#R0278 HALT protease inhibitor Thermo Fisher Cat#87786 Benzonase Millipore Cat#E1014 EDTA Thermo Fisher Cat#1861274 Tween-20 (for immunoblots) BioRad Cat#1610781 Centrifugal filter Millipore Cat#UFC30GV00 7.5% protein gel BioRad Cat#4568023 Precision Plus Protein Dual Color Standards BioRad Cat#1610374 Immobilon-P membrane Millipore Cat#IPVH00010 AP immune star substrate BioRad Cat#1705018 Experimental models: Organisms/strains Drosophila stocks used in this study, see Table S1 This study N/A Oligonucleotides Alexa 647-labeled Oligopaint probe (OPP122), Mixture of 80-base oligos targeting 100kb distal region of the X chromosome (dm6, nucleotides 1,400,000-1,500,000) Joyce Lab, University of Pennsylvania N/A (Continued on next page) Current Biology 34, 1–12.e1–e6, July 8, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cy3-conjugated probe (50-Cy3-AGGGATCGTTAGCACTCGTAAT) hybridizes to 359-bp repeat in pericentric heterochromatin of the X chromosome Integrated DNA Technologies N/A Smc3* Fwd: 5’GACAAGACGCGCCGCACGCTAG AATACATTCGGTATGAGACCGAGCTGAAGGACA Integrated DNA Technologies N/A Smc3* Rev: 5’TGTCCTTCAGCTCGGTCTCATACCG AATGTATTCTAGCGTGCGGCGCGTCTTGTC Integrated DNA Technologies N/A HA stop Fwd: 5’GCTTCTAGCGAAAAAGTGTA GACGGTATCGATAAGCTTG Integrated DNA Technologies N/A HA stop Rev: 5’CAAGCTTATCGATACCGTC TACACTTTTTCGCTAGAAGC Integrated DNA Technologies N/A Smc3-WT Fwd: 5’AGCACTGGTACCGCGGCCG CAATAAGATGCACATCAAGCAG Integrated DNA Technologies N/A Smc3-WT Rev: 5’ACTGATCTCGAGGCGGCG TGGGTGCTGTCG Integrated DNA Technologies N/A Smc3-KK Fwd: 5’CGCGTGATTGGCGCCAGAA GGGACCAGTACTTCC Integrated DNA Technologies N/A Smc3-KK Rev: 5’GGAAGTACTGGTCCCTTCTGG CGCCAATCACGCG Integrated DNA Technologies N/A Recombinant DNA Clone RE14758 (Smc3 cDNA) DGRC Stock 8520; RRID:DGRC_8502 pUASP-attB Gift from Hawley lab49 RRID: N/A w+ attB vector Addgene (donated by Jeff Sekelsky) Plasmid# 30326; RRID: Addgene_30326 Software and algorithms Volocity Visualization, Restoration, and Quantitation Quorum Technologies, Canada Version 6.5.0 https://www.volocity4d.com/ downloadhttps://www. volocity4d.com/download Volocity Acquisition (for epifluorescence imaging) Quorum Technologies, Canada Version 6.5.1 https://www.volocity4d.com/ volocity-acquisition Nikon Elements (for spinning disc confocal imaging) Nikon Version 5.11.02 Build 1369 MATLAB Mathworks Version R2022a https://www.mathworks.com/ products/new_products/ release2022a.html BioRad Image Lab BioRad Version 6.1 https://www.bio-rad.com/ en-us/product/image-labsoftware?ID=KRE6P5E8Z Microsoft Office Microsoft Version 16.84 Affinity Designer Affinity Version 1.10.6.1665 https://affinity.serif.com/ en-us/designer/ Other Chemi Doc Touch Imaging system BioRad Serial no. 732BR0120 e2 Current Biology 34, 1–12.e1–e6, July 8, 2024 Article
Protease Inhibitor:Article Title: Chromatin-associated cohesin turns over extensively and forms new cohesive linkages in Drosophila oocytes during meiotic prophase.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies High-affinity rat monoclonal anti-HA (clone 3F10) Roche Cat#11863423001 Lot no. 14223800; RRID: AB_390918 Mouse monoclonal anti-C3G (clone 1A8-1G2) Hawley Lab47 N/A Rabbit anti-CID Rogers Lab48 N/A GFP-Booster Alexa Fluor 488 (Alpaca anti-GFP) ChromTek Cat#gb2AF488; RRID: AB_2827573 RFP-Booster Alexa Fluor 568 (Alpaca anti-mCherry) ChromTek Cat#rb2AF568; RRID: AB_2827576 Affinity-purified guinea pig anti-Smc1 Bickel Lab26 N/A Rabbit anti-GFP Invitrogen Cat#A11122; RRID: AB_221569 Monoclonal mouse anti-alpha-tubulin (12G10) Developmental Studies Hybridoma Bank (deposited by J. Frankel and E.M. Nelson) Cat#AB_1157911; RRID:N/A Cy3 anti-rat (min-X mouse) Jackson ImmunoResearch Labs Cat#712-165-153; RRID: AB_2340667 Cy5 anti-rabbit Jackson ImmunoResearch Labs Cat#711-175-152; RRID: AB_2340607 Alexa 488 anti-mouse (min-X rat) Jackson ImmunoResearch Labs Cat#715-545-151; RRID: AB_2341099 Alkaline phosphatase goat anti-rat Sigma-Aldrich Cat#A8438; RRID: AB_258391 Alkaline phosphatase goat anti-mouse Invitrogen Cat#A16069; RRID: AB_2534742 Alkaline phosphatase goat anti-guinea pig Southern Biotech Cat#6090-04; RRID: AB_2796156 Alkaline phosphatase goat anti-rabbit Promega Cat#S373B; RRID: AB_430872 Chemicals, peptides, and recombinant proteins Formaldehyde Ted Pella Cat#18505 NP-40 Thermo Fisher Cat#28324 Tween-20 (for cytology) Thermo Fisher Cat#28320 BSA Fisher Scientific Cat#BP1605-100 DAPI Invitrogen Cat#D1306 Poly-L-lysine Sigma-Aldrich Cat#P8920 SlowFade Diamond Antifade Molecular Probes Cat#S36967 Prolong Gold Antifade Molecular Probes Cat#P36930 Vectashield Vibrance Vector Laboratories CatH-1700 RIPA buffer Sigma-Aldrich Cat#R0278 HALT protease inhibitor Thermo Fisher Cat#87786 Benzonase Millipore Cat#E1014 EDTA Thermo Fisher Cat#1861274 Tween-20 (for immunoblots) BioRad Cat#1610781 Centrifugal filter Millipore Cat#UFC30GV00 7.5% protein gel BioRad Cat#4568023 Precision Plus Protein Dual Color Standards BioRad Cat#1610374 Immobilon-P membrane Millipore Cat#IPVH00010 AP immune star substrate BioRad Cat#1705018 Experimental models: Organisms/strains Drosophila stocks used in this study, see Table S1 This study N/A Oligonucleotides Alexa 647-labeled Oligopaint probe (OPP122), Mixture of 80-base oligos targeting 100kb distal region of the X chromosome (dm6, nucleotides 1,400,000-1,500,000) Joyce Lab, University of Pennsylvania N/A (Continued on next page) Current Biology 34, 1–12.e1–e6, July 8, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cy3-conjugated probe (50-Cy3-AGGGATCGTTAGCACTCGTAAT) hybridizes to 359-bp repeat in pericentric heterochromatin of the X chromosome Integrated DNA Technologies N/A Smc3* Fwd: 5’GACAAGACGCGCCGCACGCTAG AATACATTCGGTATGAGACCGAGCTGAAGGACA Integrated DNA Technologies N/A Smc3* Rev: 5’TGTCCTTCAGCTCGGTCTCATACCG AATGTATTCTAGCGTGCGGCGCGTCTTGTC Integrated DNA Technologies N/A HA stop Fwd: 5’GCTTCTAGCGAAAAAGTGTA GACGGTATCGATAAGCTTG Integrated DNA Technologies N/A HA stop Rev: 5’CAAGCTTATCGATACCGTC TACACTTTTTCGCTAGAAGC Integrated DNA Technologies N/A Smc3-WT Fwd: 5’AGCACTGGTACCGCGGCCG CAATAAGATGCACATCAAGCAG Integrated DNA Technologies N/A Smc3-WT Rev: 5’ACTGATCTCGAGGCGGCG TGGGTGCTGTCG Integrated DNA Technologies N/A Smc3-KK Fwd: 5’CGCGTGATTGGCGCCAGAA GGGACCAGTACTTCC Integrated DNA Technologies N/A Smc3-KK Rev: 5’GGAAGTACTGGTCCCTTCTGG CGCCAATCACGCG Integrated DNA Technologies N/A Recombinant DNA Clone RE14758 (Smc3 cDNA) DGRC Stock 8520; RRID:DGRC_8502 pUASP-attB Gift from Hawley lab49 RRID: N/A w+ attB vector Addgene (donated by Jeff Sekelsky) Plasmid# 30326; RRID: Addgene_30326 Software and algorithms Volocity Visualization, Restoration, and Quantitation Quorum Technologies, Canada Version 6.5.0 https://www.volocity4d.com/ downloadhttps://www. volocity4d.com/download Volocity Acquisition (for epifluorescence imaging) Quorum Technologies, Canada Version 6.5.1 https://www.volocity4d.com/ volocity-acquisition Nikon Elements (for spinning disc confocal imaging) Nikon Version 5.11.02 Build 1369 MATLAB Mathworks Version R2022a https://www.mathworks.com/ products/new_products/ release2022a.html BioRad Image Lab BioRad Version 6.1 https://www.bio-rad.com/ en-us/product/image-labsoftware?ID=KRE6P5E8Z Microsoft Office Microsoft Version 16.84 Affinity Designer Affinity Version 1.10.6.1665 https://affinity.serif.com/ en-us/designer/ Other Chemi Doc Touch Imaging system BioRad Serial no. 732BR0120 e2 Current Biology 34, 1–12.e1–e6, July 8, 2024 Article
Western Blot:Article Title: Chromatin-associated cohesin turns over extensively and forms new cohesive linkages in Drosophila oocytes during meiotic prophase.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies High-affinity rat monoclonal anti-HA (clone 3F10) Roche Cat#11863423001 Lot no. 14223800; RRID: AB_390918 Mouse monoclonal anti-C3G (clone 1A8-1G2) Hawley Lab47 N/A Rabbit anti-CID Rogers Lab48 N/A GFP-Booster Alexa Fluor 488 (Alpaca anti-GFP) ChromTek Cat#gb2AF488; RRID: AB_2827573 RFP-Booster Alexa Fluor 568 (Alpaca anti-mCherry) ChromTek Cat#rb2AF568; RRID: AB_2827576 Affinity-purified guinea pig anti-Smc1 Bickel Lab26 N/A Rabbit anti-GFP Invitrogen Cat#A11122; RRID: AB_221569 Monoclonal mouse anti-alpha-tubulin (12G10) Developmental Studies Hybridoma Bank (deposited by J. Frankel and E.M. Nelson) Cat#AB_1157911; RRID:N/A Cy3 anti-rat (min-X mouse) Jackson ImmunoResearch Labs Cat#712-165-153; RRID: AB_2340667 Cy5 anti-rabbit Jackson ImmunoResearch Labs Cat#711-175-152; RRID: AB_2340607 Alexa 488 anti-mouse (min-X rat) Jackson ImmunoResearch Labs Cat#715-545-151; RRID: AB_2341099 Alkaline phosphatase goat anti-rat Sigma-Aldrich Cat#A8438; RRID: AB_258391 Alkaline phosphatase goat anti-mouse Invitrogen Cat#A16069; RRID: AB_2534742 Alkaline phosphatase goat anti-guinea pig Southern Biotech Cat#6090-04; RRID: AB_2796156 Alkaline phosphatase goat anti-rabbit Promega Cat#S373B; RRID: AB_430872 Chemicals, peptides, and recombinant proteins Formaldehyde Ted Pella Cat#18505 NP-40 Thermo Fisher Cat#28324 Tween-20 (for cytology) Thermo Fisher Cat#28320 BSA Fisher Scientific Cat#BP1605-100 DAPI Invitrogen Cat#D1306 Poly-L-lysine Sigma-Aldrich Cat#P8920 SlowFade Diamond Antifade Molecular Probes Cat#S36967 Prolong Gold Antifade Molecular Probes Cat#P36930 Vectashield Vibrance Vector Laboratories CatH-1700 RIPA buffer Sigma-Aldrich Cat#R0278 HALT protease inhibitor Thermo Fisher Cat#87786 Benzonase Millipore Cat#E1014 EDTA Thermo Fisher Cat#1861274 Tween-20 (for immunoblots) BioRad Cat#1610781 Centrifugal filter Millipore Cat#UFC30GV00 7.5% protein gel BioRad Cat#4568023 Precision Plus Protein Dual Color Standards BioRad Cat#1610374 Immobilon-P membrane Millipore Cat#IPVH00010 AP immune star substrate BioRad Cat#1705018 Experimental models: Organisms/strains Drosophila stocks used in this study, see Table S1 This study N/A Oligonucleotides Alexa 647-labeled Oligopaint probe (OPP122), Mixture of 80-base oligos targeting 100kb distal region of the X chromosome (dm6, nucleotides 1,400,000-1,500,000) Joyce Lab, University of Pennsylvania N/A (Continued on next page) Current Biology 34, 1–12.e1–e6, July 8, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cy3-conjugated probe (50-Cy3-AGGGATCGTTAGCACTCGTAAT) hybridizes to 359-bp repeat in pericentric heterochromatin of the X chromosome Integrated DNA Technologies N/A Smc3* Fwd: 5’GACAAGACGCGCCGCACGCTAG AATACATTCGGTATGAGACCGAGCTGAAGGACA Integrated DNA Technologies N/A Smc3* Rev: 5’TGTCCTTCAGCTCGGTCTCATACCG AATGTATTCTAGCGTGCGGCGCGTCTTGTC Integrated DNA Technologies N/A HA stop Fwd: 5’GCTTCTAGCGAAAAAGTGTA GACGGTATCGATAAGCTTG Integrated DNA Technologies N/A HA stop Rev: 5’CAAGCTTATCGATACCGTC TACACTTTTTCGCTAGAAGC Integrated DNA Technologies N/A Smc3-WT Fwd: 5’AGCACTGGTACCGCGGCCG CAATAAGATGCACATCAAGCAG Integrated DNA Technologies N/A Smc3-WT Rev: 5’ACTGATCTCGAGGCGGCG TGGGTGCTGTCG Integrated DNA Technologies N/A Smc3-KK Fwd: 5’CGCGTGATTGGCGCCAGAA GGGACCAGTACTTCC Integrated DNA Technologies N/A Smc3-KK Rev: 5’GGAAGTACTGGTCCCTTCTGG CGCCAATCACGCG Integrated DNA Technologies N/A Recombinant DNA Clone RE14758 (Smc3 cDNA) DGRC Stock 8520; RRID:DGRC_8502 pUASP-attB Gift from Hawley lab49 RRID: N/A w+ attB vector Addgene (donated by Jeff Sekelsky) Plasmid# 30326; RRID: Addgene_30326 Software and algorithms Volocity Visualization, Restoration, and Quantitation Quorum Technologies, Canada Version 6.5.0 https://www.volocity4d.com/ downloadhttps://www. volocity4d.com/download Volocity Acquisition (for epifluorescence imaging) Quorum Technologies, Canada Version 6.5.1 https://www.volocity4d.com/ volocity-acquisition Nikon Elements (for spinning disc confocal imaging) Nikon Version 5.11.02 Build 1369 MATLAB Mathworks Version R2022a https://www.mathworks.com/ products/new_products/ release2022a.html BioRad Image Lab BioRad Version 6.1 https://www.bio-rad.com/ en-us/product/image-labsoftware?ID=KRE6P5E8Z Microsoft Office Microsoft Version 16.84 Affinity Designer Affinity Version 1.10.6.1665 https://affinity.serif.com/ en-us/designer/ Other Chemi Doc Touch Imaging system BioRad Serial no. 732BR0120 e2 Current Biology 34, 1–12.e1–e6, July 8, 2024 Article
Membrane:Article Title: Chromatin-associated cohesin turns over extensively and forms new cohesive linkages in Drosophila oocytes during meiotic prophase.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies High-affinity rat monoclonal anti-HA (clone 3F10) Roche Cat#11863423001 Lot no. 14223800; RRID: AB_390918 Mouse monoclonal anti-C3G (clone 1A8-1G2) Hawley Lab47 N/A Rabbit anti-CID Rogers Lab48 N/A GFP-Booster Alexa Fluor 488 (Alpaca anti-GFP) ChromTek Cat#gb2AF488; RRID: AB_2827573 RFP-Booster Alexa Fluor 568 (Alpaca anti-mCherry) ChromTek Cat#rb2AF568; RRID: AB_2827576 Affinity-purified guinea pig anti-Smc1 Bickel Lab26 N/A Rabbit anti-GFP Invitrogen Cat#A11122; RRID: AB_221569 Monoclonal mouse anti-alpha-tubulin (12G10) Developmental Studies Hybridoma Bank (deposited by J. Frankel and E.M. Nelson) Cat#AB_1157911; RRID:N/A Cy3 anti-rat (min-X mouse) Jackson ImmunoResearch Labs Cat#712-165-153; RRID: AB_2340667 Cy5 anti-rabbit Jackson ImmunoResearch Labs Cat#711-175-152; RRID: AB_2340607 Alexa 488 anti-mouse (min-X rat) Jackson ImmunoResearch Labs Cat#715-545-151; RRID: AB_2341099 Alkaline phosphatase goat anti-rat Sigma-Aldrich Cat#A8438; RRID: AB_258391 Alkaline phosphatase goat anti-mouse Invitrogen Cat#A16069; RRID: AB_2534742 Alkaline phosphatase goat anti-guinea pig Southern Biotech Cat#6090-04; RRID: AB_2796156 Alkaline phosphatase goat anti-rabbit Promega Cat#S373B; RRID: AB_430872 Chemicals, peptides, and recombinant proteins Formaldehyde Ted Pella Cat#18505 NP-40 Thermo Fisher Cat#28324 Tween-20 (for cytology) Thermo Fisher Cat#28320 BSA Fisher Scientific Cat#BP1605-100 DAPI Invitrogen Cat#D1306 Poly-L-lysine Sigma-Aldrich Cat#P8920 SlowFade Diamond Antifade Molecular Probes Cat#S36967 Prolong Gold Antifade Molecular Probes Cat#P36930 Vectashield Vibrance Vector Laboratories CatH-1700 RIPA buffer Sigma-Aldrich Cat#R0278 HALT protease inhibitor Thermo Fisher Cat#87786 Benzonase Millipore Cat#E1014 EDTA Thermo Fisher Cat#1861274 Tween-20 (for immunoblots) BioRad Cat#1610781 Centrifugal filter Millipore Cat#UFC30GV00 7.5% protein gel BioRad Cat#4568023 Precision Plus Protein Dual Color Standards BioRad Cat#1610374 Immobilon-P membrane Millipore Cat#IPVH00010 AP immune star substrate BioRad Cat#1705018 Experimental models: Organisms/strains Drosophila stocks used in this study, see Table S1 This study N/A Oligonucleotides Alexa 647-labeled Oligopaint probe (OPP122), Mixture of 80-base oligos targeting 100kb distal region of the X chromosome (dm6, nucleotides 1,400,000-1,500,000) Joyce Lab, University of Pennsylvania N/A (Continued on next page) Current Biology 34, 1–12.e1–e6, July 8, 2024 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Cy3-conjugated probe (50-Cy3-AGGGATCGTTAGCACTCGTAAT) hybridizes to 359-bp repeat in pericentric heterochromatin of the X chromosome Integrated DNA Technologies N/A Smc3* Fwd: 5’GACAAGACGCGCCGCACGCTAG AATACATTCGGTATGAGACCGAGCTGAAGGACA Integrated DNA Technologies N/A Smc3* Rev: 5’TGTCCTTCAGCTCGGTCTCATACCG AATGTATTCTAGCGTGCGGCGCGTCTTGTC Integrated DNA Technologies N/A HA stop Fwd: 5’GCTTCTAGCGAAAAAGTGTA GACGGTATCGATAAGCTTG Integrated DNA Technologies N/A HA stop Rev: 5’CAAGCTTATCGATACCGTC TACACTTTTTCGCTAGAAGC Integrated DNA Technologies N/A Smc3-WT Fwd: 5’AGCACTGGTACCGCGGCCG CAATAAGATGCACATCAAGCAG Integrated DNA Technologies N/A Smc3-WT Rev: 5’ACTGATCTCGAGGCGGCG TGGGTGCTGTCG Integrated DNA Technologies N/A Smc3-KK Fwd: 5’CGCGTGATTGGCGCCAGAA GGGACCAGTACTTCC Integrated DNA Technologies N/A Smc3-KK Rev: 5’GGAAGTACTGGTCCCTTCTGG CGCCAATCACGCG Integrated DNA Technologies N/A Recombinant DNA Clone RE14758 (Smc3 cDNA) DGRC Stock 8520; RRID:DGRC_8502 pUASP-attB Gift from Hawley lab49 RRID: N/A w+ attB vector Addgene (donated by Jeff Sekelsky) Plasmid# 30326; RRID: Addgene_30326 Software and algorithms Volocity Visualization, Restoration, and Quantitation Quorum Technologies, Canada Version 6.5.0 https://www.volocity4d.com/ downloadhttps://www. volocity4d.com/download Volocity Acquisition (for epifluorescence imaging) Quorum Technologies, Canada Version 6.5.1 https://www.volocity4d.com/ volocity-acquisition Nikon Elements (for spinning disc confocal imaging) Nikon Version 5.11.02 Build 1369 MATLAB Mathworks Version R2022a https://www.mathworks.com/ products/new_products/ release2022a.html BioRad Image Lab BioRad Version 6.1 https://www.bio-rad.com/ en-us/product/image-labsoftware?ID=KRE6P5E8Z Microsoft Office Microsoft Version 16.84 Affinity Designer Affinity Version 1.10.6.1665 https://affinity.serif.com/ en-us/designer/ Other Chemi Doc Touch Imaging system BioRad Serial no. 732BR0120 e2 Current Biology 34, 1–12.e1–e6, July 8, 2024 Article
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