How do you calculate DNA Picomoles?
How do you calculate DNA Picomoles?
Picomoles/µl = DNA Concentration(µg/ml) / [0.66*DNA Size(bp)]
How do you calculate molecular weight from blood pressure?
The molecular weight or molar mass of any double-stranded DNA fragment can therefore be calculated by multiplying its length (in bp) by 650 and the answer will be expressed as daltons or g/mol.
How do you calculate molarity of plasmid?
If your two plasmids have similar proportions of each base then the ratio of molecular weights will be 1: 1.433 (A:B) so for a one to one molar mix in 1.5ug you need to add 1.433/2.433 x 1.5ug = 0.883ug of B and 1/2.433 x 1.5ug = 0.617ug of A.
What is ligation ratio?
Keep total DNA concentration between 1-10 µg/ml. Vector: Insert molar ratios between 1:1 and 1:10 are optimal for single insertions (up to 1:20 for short adaptors). Insert: vector molar ratio should be 6:1 to promote multiple inserts.
How do you convert nanograms to Picomoles?
Given here unit conversion formula to convert the given values of Pmol/L to Ng/Ml. pmol/L to ng/ML conversion formula is derived as n = p / 2.247 where, n = ng/mL, p = pmol/L. Ng/ml stands for nanograms per milliliter whereas Pmol/L stands for Picomoles per Liter.
Is pmol UL the same as uM?
pmol/uL↔uM 1 pmol/uL = 1 uM.
How do you calculate molecular weight from gel electrophoresis?
Use a graphing program, plot the log (MW) as a function of Rf. Generate the equation y = mx + b, and solve for y to determine the MW of the unknown protein. Run the standards and samples on an SDS-PAGE gel. Process the gel with the desired stain and then destain to visualize the protein bands.
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 much DNA do I need for my ligation reaction?
However, for most standard cloning (where the insert is smaller than the vector) a 3 insert : 1 vector molar ratio will work just fine. We recommend around 100ng of total DNA in a standard ligation reaction. Use a ligation calculator to easily quantify how much vector and insert DNA to use. Combine the following in a PCR or Eppendorf tube:
Where can I find a ligation calculator for my project?
Ligation calculators are easily found on the web. Just enter the concentration, lengths of your insert and vector, and what ratio you want, and it will tell you exactly how much of each to use.
What is the standard protocol for DNA ligation?
Protocol: Standard Insert + Vector DNA Ligation Before setting up the ligation reaction itself, it is important to determine the amount of cut insert and vector to use for the ligation reaction. The volume of vector DNA and insert DNA used in the ligation will vary depending on the size of each and their concentration.
How do I ligation my pGem®-T and pGem-T easy vector systems?
Alternatively, a double-digestion may be used to release the insert from either vector. Rapid Ligation:The pGEM®-T and pGEM -T Easy Vector Systems are supplied with 2X Rapid Ligation Buffer. Ligation reactions using this buffer may be incubated for 1 hour at room temperature.
