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[DOC File]Electro-competent
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Transformation Protocol. 1. Thaw competent cells on ice. It is essential that cells be completely thawed before pipetting. Pipet slowly. 2. Transfer 60 ul cells to a prechilled Falcon tube, then add DNA of your choice (pure DNA or ligation mix) to the cells and incubate on …
e.coli competent cell protocol

Competent Cell Preparation | Şen Lab
KCM Competent Transformation Protocol. Thaw an aliquot of KCM competent cells on ice. Remove 100 uL from the aliquot and place it in a fresh tube. Add 30 uL 5x KCM solution and 50 uL ddH20 to the cells. Add your DNA (1 uL of prepped plasmid is more than sufficient). Incubate cells on ice for 5 minutes. Heat shock cells at 42 C for 1-2 minutes.
make chemically competent cells

[DOC File]KCM Competent Transformation Protocol
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TSS Competent Cells (for KCM Transformations) Day 1 Restreak the bacteria on LB plates with appropriate antibiotics from a -80 stock. It is best not to restreak from a competent cell stock. Day 2. 1. Grow up a single colony in 10mL of LB+antibiotic. 2. Autoclave large centrifuge bottles and hundreds of eppendorf tubes in covered beaker(s).
making competent cells protocol

[DOC File]DH5 alpha Competent Cells - Brigham Young University
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Place the competent cells on LB plate, 37°C overnight (16-20h). Pick a single bacterial colony from a plate that has been incubated for 16-20 hours at 37°C. Transfer the colony into 20 ml of LB medium in a 100-ml flask. Incubate the culture overnight at 37°C with vigorous aeration (250 rpm in a rotary shaker).
dh5 alpha competent cells protocol

[DOC File]Heat shock transformation with frozen competent cells
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Get the ice bucket and keep DH5a chemical competent cells (from the -80癈e freezer) in ice for 5~10 minutes for them to thaw. 2. Turn on the heat shock machine to low setting at 42°C. Add water to the surface to make a water bath for even heat-up. 3. Put LB plate with the appropriate antibiotics at 37癈f incubator to warm up. ...
preparation of competent cells

Ultra-high Efficiency DH5 Competent Cells
When needed, remove a tube of competent cells from the -70ºC freezer. Thaw the cells by holding the tube in the palm of the hand. Just as the cells thaw, transfer the tube to an ice bath. Store the cells on ice for 10 minutes. Use a chilled, sterile pipette tip to transfer the competent cells to chilled, sterile 17 x 100-mm polypropylene tubes.
chemically competent cells protocol

[DOC File]TSS Competent Cells (for KCM Transformations)
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6. Pellet the cells by centrifugation at 4,500 x g for 5 minutes at 4°C. 7. Gently resuspend the cells in 1/25 of the original culture volume of ice-cold TFB2. For a 250 ml subculture, use 10 ml of TFB2. Note: Treat the competent cells gently as they are highly sensitive to handling and elevated temperature. 8.
super competent cell protocol

[DOC File]The Inoue Method for Preparation and Transformation of ...
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-80°C Frozen Heat Shock (HS) competent E. coli cells (400 L aliquots) Water bath or heat block (with water) at 42°C – check for this before you start! LB broth without selection and LB plates with appropriate selection. Collect an aliquot of HS competent E. coli from the -80°C freezer and thaw on ice.
how to make cells competent

Ultra-high Efficiency DH5 Competent Cells
It is recommended that competent cells be used within 3 months after arrival. Transformation Protocol. 1. Thaw competent cells on ice. It is essential that cells be completely thawed before pipetting. Pipet slowly. 2. Add DNA of your choice (pure DNA or ligation mix) to the cells and incubate on ice for 30 minutes (may use 5 min for subcloning).
e.coli competent cell protocol

[DOC File]Competent cell preparation¬ using rubidium chloride
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Groselab Protocol for Preparation of DH5α . Chemically Competent Cells. It is easiest to make a whole bunch of competent cells at once. They may be stability kept at -80˚C for up to a year or more. Day 1. Streak DH5α for isolation on LB plate (no ampicillin).
make chemically competent cells

Competent cells protocol