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

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USD$135.00
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Quick Overview

Download: MSDS PDF | USER MANUAL PDF


SLIC-Easy chemical competent cell preparation kit (SLIC-1001) allows user to produce about 30 vials of chemically competent E.Coli cells suitable for SLIC reaction in just one day.


Expected transformation efficiency 108~ 109 cfu/ µg DNA


Kit components:  store at 4 °C


Lyophilized SLIC-Easy E.Coli in 2 ml tube (Cat# SLIC1001-01)


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


Storage solution: 2.0ml in 2 ml tube (Cat# SLIC-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- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(StrR) endA1.

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

Details

SLIC-Easy™ chemical competent cell preparation kit (SLIC-1001) allows user to produce about 30 vials of chemically competent E.Coli cells suitable for amplification of viral plasmid in just one day. Expected transformation efficiency 108~ 10cfu/ µg DNA.

Procedure:  Estimated time 5-7 hrs.

1. The day before.  Reconstitute lyophilized SLIC-EasyTME.Coli with 2ml LB medium mixture in a 14 ml sterile tube such as FALCON 352059. Incubate at 37 °C 220 rpm.

Add 25 µg/ml streptomycin after 1 hr (optional). Grow overnight.

2. Next morning. Prepare LB medium mixture:

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

3. Add 0.3 ml overnight culture to 40 ml LB medium prepared.

4. Incubate the medium at 37°C 220 rpm shaker till OD 600nm becomes between 0.3-0.5 (below 0.7 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 SLIC-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.

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