What is a 1 kb DNA ladder?
1 kb DNA Ladder allows for determining the size of double-stranded DNA from 250 – 10,000 base pairs (bp). The ladder consists of 13 double-stranded, blunt-end fragments, of sizes 250, 500, 750, 1000, 1500, 2000, 2500, 3000, 4000, 5000, 6000, 8000, and 10,000 bp (see Figure 1).
Do you add loading dye to DNA ladder?
This protocol is recommended for a 5mm wide gel lane….Dilute only 1 µl of DNA Ladder at a time.
| Distilled water (dH20)* or TE Buffer | 4 μl |
|---|---|
| Gel Loading Dye, Purple (6X), no SDS | 1 μl |
| DNA Ladder/Marker | 1 μl |
| Total volume | 6 μl |
Can I dilute DNA ladder?
Dilute only 1 µl of DNA Ladder at a time DNA may denature if diluted and stored in dH20. This protocol is recommended for a 5mm wide gel lane. The components of the mixture should be scaled up or down, depending on the width of the lane.
What is bp in gel electrophoresis?
The term ‘bp’ in gel electrophoresis stands for base pair. Base pairs refer to a set of complementary nucleotides in DNA.
Why do gels smile?
The “smile” effect where the center lanes run faster than the outside lanes is common with many gel boxes. One way to reduce that is to run the gel at a lower voltage for a longer time. The “curling” of individual bands is usually a result of overloading the wells (too much DNA).
Why is the DNA ladder not separating?
When increasing percentage of low melting agarose gel (1.5% low melting or 1% low melting) the bands and ladder do not separate enough, when running on lower voltage such as 60-80V, for 2-3hours. Increasing voltage would melt the low-melting agarose.
What is 100bp ladder?
100 bp DNA Ladder consists of 13 individual chromatography-purified DNA fragments and has reference bands at 2000, 1500, and 600 bp for easy orientation. 100 bp DNA Ladder is ideal for separation on 1–2% agarose gels.