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Omni-Easy™ Chemical Competent Cell Kit (Omni-1001)

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USD$120.00
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Quick Overview

Download: MSDS PDF


Omni-Easychemical competent cell preparation kit (Omni-1001) allows user to produce about 30 vials of chemically competent E.Colicells suitable for general purpose subcloning and amplification, especially for both SLIC reaction and virus plasmids amplification. It is not suitable for blue/white screening.


Expected transformation efficiency 108~ 109 cfu/ µg DNA


Kit components:  store at 4 °C


Lyophilized Omni-Easy E.Coli in 2 ml tube (Cat# Omni-1001-01)


LB Supplement: 0.5ml in 2 ml tube (Cat# Omni-LB-500)           


Storage solution: 2.0ml in 2 ml tube (Cat# Omni-STOR-2000)


User prepared materials and equipment:


1. LB medium (40ml).


2. Centrifuge capable to hold 50ml tube 1500 x g at 4 °C


3. Tube to store competent cells.


E.Coli information


Genotype: F–, thi-1, hsdS20 (rB, mB), supE44, recA13, ara-14, leuB6, proA2, lacY1, galK2, rpsL20 (strr), xyl-5, mtl-1

Omni-Easy™ Chemical Competent Cell Kit (Omni-1001)

Details

Omni-Easy chemical competent cell preparation kit (Omni-1001) allows user to produce about 30 vials of chemically competent E.Coli cells suitable for general purpose subcloning and amplification, especially for both SLIC reaction and virus plasmids amplification. It is not suitable for blue/white screening.

Procedure:  Estimated time 5-7 hrs.

1. Prepare LB medium mixture:

  • 50ml tube containing autoclaved 40ml LB medium.
  •  Add 0.4ml LB Supplement to the LB medium.

2. Reconstitute lyophilized Omni-Easy E.Coli with 1ml LB medium mixture.

3. Take it to 40ml LB medium mixture.

4. Incubate the medium at 37°C 220 rpm shaker till OD 600nm becomes between 0.3-0.4 (below 0.5 is still acceptable).

To get highest transformation efficiency, following procedure should be performed in cold room (4°C). 

Prepare the storage box in cold room.

5.  Put the tube into ice for 10min.

6. Centrifuge 50ml tube containing Omni-Easy E.Coli at 1500 x g  for 10min at 4°C.

7. Carefully decant supernatant and leave the tube upside-down for 2-3min.

8. Resuspend the E.Coli pellet with 1ml storage solution by using a sterile glass rod or Pasteur pipet (Do not vortex or pipet up-down).

9. Add another 1ml of storage solution. Keep on ice another 20min.

10. Aliquot out 65µl each into a screw-capped small sterile micro-tube such as MBP3469. 

11. Put into the storage box and store at - 80°C.

Expected transformation efficiency 108~ 109 cfu/ µg DNA

Usage

1. Thaw the tube on ice. 

2. Mix with plasmid (up to 4 µl) and incubate on ice for 10-30 min.

3. Heat shock at 42 °C for 60 second.

4. Chill on ice for 1- 2 min.

5. Add 300 µl SOC medium.

6. Incubate at 37°C for 1hr.

7. Plate on appropriate antibiotics agar. 

Related Products

VP-Easy Chemical Competent Cell Preparation Kit (VP1001)

SLIC-Easy Chemical Competent Cell Preparation Kit (SLIC-1001)


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