How do you calculate copy number of plasmid?

How do you calculate copy number of plasmid?

Plasmid copy number was determined by comparing the quantification signal from the plasmid to those from the chromosome. Copy number was then calculated by using a known copy number plasmid as a standard.

How do you calculate ng of DNA?

  1. ng is the amount of DNA (plasmid, primer, etc.) you have in nanograms.
  2. 6.022 x 1023 = Avogadro’s number.
  3. Length is the length of your DNA fragment in base pairs. Just multiply by 1000 if you are working in kb.
  4. We multiply by 1 x 109 to convert our answer to nanograms.

How many Ng is a plasmid?

5-10 ng of plasmid/cosmid DNA are required. 1-For transformation, you could use very less DNA, 1 ng.

How do I know if my plasmid is a high or low copy number type?

A way to determine experimentally if the copy number of your plasmid is high or low is to perform a miniprep. A high-copy plasmid should yield between 3-5 ug DNA per 1 ml LB culture, while a low-copy plasmid will yield between 0.2-1 ug DNA per ml of LB culture.

How do I find my transgene copy number?

Considering zygosity, the transgene copy number should be multiple times of 0.5 as compared to reference gene. In other words, a hemizygous, one-copy transgene will be 0.5 times of the reference gene. A homozygous one copy transgene will be one (2 by 0.5) times the reference gene.

How much DNA does it take to transform E coli?

For successful chemical transformation, 50–100 µL of competent cells and 1–10 ng of DNA are recommended. When a ligation mixture is used as the transforming DNA (often 1–5 µL is sufficient), purification prior to chemical transformation is generally not required.

How many FG are in a ng?

Nanogram to Femtogram Conversion Table

Nanogram [ng] Femtogram [fg]
1 ng 1000000 fg
2 ng 2000000 fg
3 ng 3000000 fg
5 ng 5000000 fg