What are one shot competent cells?
One Shot TOP10 Chemically Competent E. coli are provided at a transformation efficiency of 1 x 109 cfu/μg plasmid DNA. They allow stable replication of high-copy number plasmids and are the same competent cells that come with many Life Technologies cloning kits.
How do I make DH5 alpha competent cells?
1) Inoculate an LB culture with DH5α cells (directly from the frozen stock without thawing) and grow overnight at 37 °C. 2) Add 5 mL of this overnight culture to 500 mL of SOB medium in a 2 L flask. 3) Grow cells to an OD600 between 0.4 or 0.6. Do not exceed 0.6.
How do you make ecoli competent?
Take the strain of E. coli you wish to make chemically competent from either a glycerol stock or a freshly-streaked agar plate and inoculate it into a flask containing approximately 50 mL of RB. Let it grow overnight. As I have said, no antibiotics are needed in this step!
How does CaCl2 make cells competent?
The process of calcium chloride heat-shock transformation encourages bacterial cells to uptake DNA from the surrounding environment. The ice-cold CaCl2 solution facilitates binding of DNA to the surface of the cell, which then enters the cell after a short period of heat- shock (3).
What is TOP10 E. coli?
Product description. TOP10 E. coli are provided at a transformation efficiency of 1 × 109 cfu/μg supercoiled DNA and are ideal for high-efficiency cloning and plasmid propagation. They allow stable replication of high-copy number plasmids. The genotype of TOP10 Cells is similar to the DH10B™ strain.
Does BL21 have T7 polymerase?
BL21(DE3)pLysS is a derivative of BL21 that has the T7 RNA polymerase gene under the control of the lacUV5 promoter. This arrangement is on a phage genome, called DE3. DE3 is inserted into the chromosome of BL21 to make BL21(DE3).
When to use a support supplement for methylation?
Methylation is carried out by specific enzymes, each one using specific nutrients. Use support supplements only when a strong nutritional foundation has been established and only when needed. Supplements can assist in providing specific cofactors required in enzymatic reactions and can protect against the stress hormones.
What foods to eat to support your methylation cycle?
At the same time, choosing foods that are packed with active folate (uncooked leafy greens) and antioxidants (think colorful fruits and veggies) will support methylation and help decrease your oxidative stress.
What are the cofactors for the methylation enzymes?
Cofactors are other nutrients that the enzymes need to do their job properly. Your practitioner will be able to advise which cofactors you need. Cofactors for the methylation enzymes are: Riboflavin (B2 or R5P) Zinc. Choline. TMG (in other products) Vitamin B6 (or P5P)
What kind of vitamins are in methyl Guard plus?
Methyl-Guard Plus contains 5MTHF (the active form of folate), pyridoxal 5′-phosphate (active B6), methylcobalamin (active B12), Betaine Anhydrous (Trimethylglycine or TMG) and riboflavin 5′-phosphate (the active form of vitamin B2) for its positive effect on homocysteine metabolism and methylation.