FAQs

1. How should I prepare the DNA?
Please provide column-purified (Qiagen) plasmid DNA with the following specifications:

  • P-element services: 1–2 µg/µl concentration in 20 µl ultrapure water or Injection Buffer (IB).
  • Site-specific services: 0.75–1 µg/µl concentration in 20 µl ultrapure water or Injection Buffer (IB).

Composition for 50X IB: 0.5 mM sodium phosphate buffer (pH 7.5) and 5 mM KCl.

2. Is there a special requirement for DNA in the Site-Specific service?
No special requirement. The same quality of DNA as described above is required: 0.75–1 µg/µl of plasmid DNA in 20 µl ultrapure water or Injection Buffer (IB).

3. Which fly strains do you use for injections?
We use:

Fly Stocks we typically use for site specific Integration :

  • Bloom - 79604 - y w P{y=nanos-phiC31\int.NLS}X; P{y=CaryP}attP40 - Chr 2, 25C6, 2L:5108448 - regularly used
  • Bloom - 25709 - y v P{y=nos-phiC31\int.NLS}X; P{y=CaryP}attP40 - Chr 2, 25C6, 2L:5108448 - regularly used
  • Bloom - 25710 - P{y=nos-phiC31\int.NLS}X, y sc v sev; P{y=CaryP}attP2 - Chr 3, 68A4, 3L:11070538 - regularly used
  • Bloom - 99002 - y w P{y=nanos-phiC31\int.NLS}X; P{y=CaryP}attP2 - Chr 3, 68A4, 3L:11070538 - regularly used 

List of fly stocks available for transgenic generation 

  • Bloom - 24480 - y M{3xP3-RFP.attP}ZH-2A w; M{RFP GFP=vas-int.Dm}ZH-102D - Chr X
  • Bloom - 24868 - y M{RFP GFP=vas-int.Dm}ZH-2A w; PBac{y-attP-9A}VK00022 - Chr 2, 57F5, 2R:21645971
  • Bloom - 24867 - y M{RFP GFP=vas-int.Dm}ZH-2A w; PBac{y-attP-9A}VK00020 - 3R:30,553,313..30,553,313
  • VK02 - y w P{y=nos-phiC31\int.NLS}X; PBac{y-attP-3B}VK00002 - 2
  • Bloom - 32542 - y w P{y=nos-phiC31\int.NLS}X; PBac{y-attP-3B}VK00033 - 3
  • Bloom - 32543 - y M{RFP GFP=vas-int.Dm}ZH-2A w v P{y=nanos-phiC31\int.NLS}X; PBac{y-attP-3B}VK00033 - 3L
  • w1118 for P-Element

List of fly stocks available for CRISPR Mutagenesis

  • Bloom - 54591 - y M{w=nanos-Cas9.P}ZH-2A w - nos Cas9 on X
  • Bloom - 51323 - y M{RFP GFP=vas-Cas9}ZH-2A w/FM7c - vas-Cas9 on X
  • Bloom - 54590 - y M{w=Act5C-Cas9.P}ZH-2A w - Act5C-Cas9 on X
  • Act5C-Cas9 Lig4 - Act5C-Cas9 on X
  • Bloom - 78781 - y1 sc* v1 sev21; P{nanos-Cas9.R}attP40 - nos Cas9 on 2nd
  • Bloom - 51324 - w; PBac{y GFP=vas-Cas9}VK00027 - vas-Cas9 on 3rd

4. Which helper plasmid should I use for P-element transformation?
We prefer the Δ2-3 plasmid. Please send at least 20 µl of column-purified helper plasmid DNA, clearly labeled with the concentration. The concentration should be more than 1 µg/µl.

5. Which balancers do you use to balance the insertion?
We normally use:

  • X chromosome: FM7a
  • II chromosome: CyO
  • III chromosome: TM3Sb/TM6Tb

If needed, other balancers can be used. Charges may apply depending on the availability of the stocks.

6. How long does it take to receive transformants?
Timelines vary based on service plan and workload.

  • For the Site-Specific Excel plan, it typically takes 8–10 weeks from DNA injection to final stock shipping.
    Updates will be provided through our web portal.

7. For how long do you keep the flies after submission?
Balanced lines are retained for 2 weeks after you receive the shipment. If there are any issues, please inform us within this time so we can send backup flies.

8. How are flies shipped?

  • Within India: Courier
  • International shipments: FedEx

9. Whom should I contact for more information?
You can reach us at: