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PhiC31

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PhiC31 Integration System

 

PhiC31 integration system is a site-specific integration approach which applies bacteriophage PhiC31 integrase to mediate the sequence-specific recombination between two largely different attachment sites (attB and attP). By selection of attB or attP sites in the genome (landing platforms) and a plasmid containing the complementary site (an integration vector), transgen can be integrated on the pre-selected site on the genome.

 

We thank Dr. Basler lab, Dr. Bellen lab, Dr. Calos lab and Dr. Perkins lab for allowing our services to use the attP lines generated in their labs. We also thanks Bloomington Stock Center for providing all these lines.

Reference: 
P{CaryP} insertions:
Groth et al. 2004
PBac{y[+]-attP} insertions: Venken et al. 2006
M{3xP3-RFP.attP}, M{vas-int.B} and P{nos-phiC31\int.NLS} insertions: Bischof et al. 2007 (see FlyC31 for additional information the phiC31 integrase system)
M{vas-int.Dm} insertions: K. Basler and F. Karch,
FlyC31
P{attP.w[+].attP} insertions:  Bateman et al., 2006

We provide micro-injection service for almost all the phiC31 lines listed in Bloomington Stock Center (table below). The service includes injection only service (service A) and extended service for screening the transformants (service B).

 

Service A: injection of 150 to 200 embryos of selected line, sending back 50 to 100 survival larvae with service report.

 

Service B: including service A, plus setting up 30 to 50 crosses, sending back transformants with service report.

 

The price for service A: $180 plus $25 handling charge per order. If you need inject into the lines other than our collection, please provide two bottles of flies. You will be charged for extra $25 fly handling fee per order.

 

The price for service B: $280 plus $25 handling charge per order.

 

The charge for all the service is non-refundable. However, if there is no transformants, we provide one free re-injection.

 

The quality of DNA is critical for successful transformation. We recommend using Invitrogen Purelink Hipure Plasmid Filter Maxiprep kit for large DNA (size over 20kb) preparation, and Qiagen Midi or Maxi kit for size less than 15kb DNA preparation. Dissolve DNA in DEPC treated water with concentration over 1.0ug/ul.

 

RTF standard attP lines collection 

Bloomington Stock # Symbol Cytology RTF order number Survival rate Transformants
24480 M{vas-int.Dm}ZH-102D, M{3xP3-RFP.attP}ZH-2A 102D, 2A3 24480 or 2A 30%-50% +++
24481 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-22A 2A3, 22A2 24481 or 22A 30%-50% +++
24482 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-51C 2A3, 51C1 24482 or 51C 30%-50% +++
24483 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP}ZH-51D 2A3, 51D9 24483 or 51D 30%-50% +++
24484 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-58A 2A3, 58A3 24484 or 58A 30%-50% +++
24485 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-68E 2A3, 68E1 24485 or 68E 30%-50% +++
24486 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-86Fa 2A3, 86E18 24486 or 86Fa 30%-50% +++
23648** M{vas-int.B}ZH-102D, M{3xP3-RFP.attP}ZH-86Fb 102D, 86F8 23648** 30%-50% +++
24487 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP'}ZH-96E 2A3, 96E10 24487 or 96E 30%-50% +++
24488 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP}ZH-102D 2A3, 102D 24488 or 102D 30%-50% +++
23647** M{vas-int.B}ZH-86Fb, M{3xP3-RFP.attP}ZH-102D 86F8, 102D 23647** 30%-50% +++
24749 M{vas-int.Dm}ZH-2A, M{3xP3-RFP.attP}ZH-86Fb 2A3, 86F8 24749 or 86Fb 30%-50% +++
24861 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00001 2A3, 59D3 24861 9%  
24862 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00005 2A3, 75A10 24862 62% +++
24863 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00007 2A3, 82A1 24863 33%  
24865 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00016 2A3, 47C6 24865 17%  
24866 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00019 2A3, 68D2 24866 17% +++
24867 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00020 2A3, 99F8 24867 69%  
24869 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00030 2A3, 50E1 24869 23%  
24870 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00031 2A3, 62E1 24870 28% +++
24871 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00033 2A3, 65B2 24871 69% +++
24872 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-3B}VK00037 2A3, 22A3 24872 41% +++
24864 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00013 2A3, 76A2 24864 22%  
24868 M{vas-int.Dm}ZH-2A, PBac{y[+]-attP-9A}VK00022 2A3, 57F5 24868 41%  
25709 P{nos-phiC31\int.NLS}X, P{CaryP}attP40 X, 25C7 25709 72%  
25710 P{nos-phiC31\int.NLS}X, P{CaryP}attP2 X, 68A4 25710 39%  
8621 P{CaryP}attP1  55C2-4 8621 66% +++
8622 P{CaryP}attP2 68A4 8622 62% +++
9729 PBac{y[+]-attP-3B}VK00010 ch. 2 9729 62%  
9734 PBac{y[+]-attP-9A}VK00015 ch. 2 9734 28%  
9753 PBac{y[+]-attP-3B}VK00038 5B8 9753 50% +++
9726 PBac{y[+]-attP-9A}VK00006 1.90E+08 9726 48% +++
9752 PBac{y[+]-attP-3B}VK00037 22A3 9752 65% +++
9731 PBac{y[+]-attP-9A}VK00012 25A3 9731 54%  
9723 PBac{y[+]-attP-3B}VK00002 2.80E+08 9723 44%  
9724 PBac{y[+]-attP-3B}VK00003a 31B1 9724 47% +++
9730 PBac{y[+]-attP-3B}VK00011 4.00E+05 9730 37%  
9733 PBac{y[+]-attP-9A}VK00014 43A1 9733 25%  
9735 PBac{y[+]-attP-3B}VK00016 47C6 9735 41%  
9728 PBac{y[+]-attP-3B}VK00008 50A3 9728 46%  
9747 PBac{y[+]-attP-9A}VK00030 5.00E+02 9747 61% +++
9736 PBac{y[+]-attP-9A}VK00018 53B2 9736 73%  
9740 PBac{y[+]-attP-9A}VK00022 57F5 9740 42%  
9722 PBac{y[+]-attP-3B}VK00001 59D3 9722 61% +++
9748 PBac{y[+]-attP-3B}VK00031 6.20E+02 9748 53% +++
9750 PBac{y[+]-attP-3B}VK00033 65B2 9750 69% +++
9754 PBac{y[+]-attP-3B}VK00039 6.70E+05 9754 49%  
9737 PBac{y[+]-attP-9A}VK00019 68D2 9737 58% +++
9741 PBac{y[+]-attP-9A}VK00023 70A2 9741 59%  
9751 PBac{y[+]-attP-3B}VK00036 70C4 9751 46% +++
9725 PBac{y[+]-attP-9A}VK00005 75A10 9725 58% +++
9732 PBac{y[+]-attP-9A}VK00013 76A2 9732 68% +++
9739 PBac{y[+]-attP-9A}VK00021 76C5 9739 65% +++
9746 PBac{y[+]-attP-9A}VK00029 h47-h53 9746 68%  
9727 PBac{y[+]-attP-3B}VK00007 82A1 9727 38%  
9749 PBac{y[+]-attP-3B}VK00032 85A2 9749 28%  
9755 PBac{y[+]-attP-3B}VK00040 87B10 9755 68% +++
9744 PBac{y[+]-attP-9A}VK00027 8.90E+12 9744 58% +++
9745 PBac{y[+]-attP-9A}VK00028 92F1 9745 31%  
9743 PBac{y[+]-attP-9A}VK00026 96F3 9743 66%  
9738 PBac{y[+]-attP-9A}VK00020 99F8 9738 73% +++
9742 PBac{y[+]-attP-9A}VK00024 101F1 9742 78%  
27387 P{attP.w[+].attP}JB37B 37B7 27387 51%  
27388 P{attP.w[+].attP}JB38F 38F1 27388 90%  
27386 P{attP.w[+].attP}JB53F 53F8 27386 86%  
25091 P{attP.w[+].attP}tara[JB89B] 89B8 25091 60%  

Note: +++ indicates at least one transgenic line obtained in the test; ** lines no longer in use.

 

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