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VP-Easy™ Chemical Competent Cell Preparation Kit (VP1001)

Availability: In stock

USD$135.00
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Quick Overview

Download: USER MANUAL | MSDS PDF


VP-Easy™ Chemical Competent Cell Preparation Kit allows user to produce chemical competent E.Coli cells suitable for viral plasmid amplification in just one day.


Expected transformation efficiency 107cfu/µg DNA.


Kit components:  store at 4°C


Lyophilized VP-Easy™ E.Coli in 2ml tube (Cat#VP1001-01)


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


Storage solution: 2.0ml in 2ml tube (Cat#VP-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- glnV44 recA13 mcrB mrr hsdS20(rB-, mB-) ara-14 galK2 lacY1 proA2 rpsL20 xyl-5 leu.

VP-Easy™ Chemical Competent Cell Preparation Kit (VP1001)

Details

 VP-Easy™ chemical competent cell preparation kit (VP-1001) allows user to produce about 30 vials of chemically competent E.Coli cells suitable for amplification of viral plasmid in just one day. Viral plasmid in VP-Easy is more stable (less mutant) than in SLIC-Easy™ E.Coli (SLIC-1001), although transformation efficiency is not as high as SLIC-Easy. It is suitable to amplify unstable plasmid especially in a large scale preparation.

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 VP-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 VP-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 107cfu/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

SLIC-Easy™ chemical competent cell preparation kit (SLIC-1001)

Omni-EasyChemical Competent Cell  Kit (Omni-1001)

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