DH5a competent cells

DH5a competent cells

Product Name: DH5a Chemically Competitive Cell

Product Code: G6016

Storage conditions: -80 ° C

Item number, quantity, and specifications: G6016-20 (20 units 100u1), G6016-100 (100 units 100u1)|

pUC19 ( control vector) 10pg/ μ 1; 10μ 1

genotype

F-φ80 lac Z△M15△(lacZYA-arg F) U169 endAl recAl

hsdR17 (rk- , mk+) supE44 λ- thi -1 gyrA96 relAl phoA

Operating instructions|

1. CLBioDH5a competent cellsPlace in ice to melt (or let go of the heart or room temperature for a moment, and quickly insert the bacterial cells into the ice when they are in a mixed state of ice and water), add the target DNA (plasmid or ligation product) and hit it by handGently mix the bottom of the EP tube and let it stand on ice for 25 minutes.

Heat shock in a 2.42 ° C water bath for 45 seconds, quickly place it back on ice and let it stand for 2 minutes. Shaking will reduce the conversion efficiency.

3. Add 700 μ l of antibiotic free sterile culture medium (2YT or LB) to the centrifuge tube and mix well. Resuscitate at 37 ° C and 200 rpm for 60 minutes.

Centrifuge at 4.5000rpm for one minute to collect the bacteria, and take about 100 μ 1 of the supernatant. Gently blow and resuspend the bacterial blocks and coat them onto 2YT or LB medium containing the corresponding antibiotics.

5. Invert the plate and incubate overnight in a 37 ° C incubator. If performing blue and white spot screening, incubate the plate at 37 ° C for at least 17 hours.

Precautions

CLBio competent cellssuggestionMelting on ice. Leave it on ice for about 5 minutes to melt, and add the plasmid within 8 minutes after melting, otherwise it will affect the transformation efficiency.

2. When mixing plasmids or ligation products, gentle handling should be performed.

3. Converting high concentration plasmids or efficient ligation products can correspondingly reduce the amount of bacteria ultimately used for plate coating.