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You Need To Get Rid Of Your PCr Gels Troubleshooting Problems

Resolve Common PC Errors

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    You may come across an error code indicating that pcr gels will be fixed. There are several ways to solve this problem, and we will deal with it shortly. No ribbons and frosty front.The sample does not sink to the bottom of the well.The sample comes out of the well.The ribbons are spread vertically.There are too many groups.The gel flows abnormally slowly.The gel works abnormally quickly.Protein bands too close together (not fully resolved)

    Possible causes Recommendations
    Preparing the gel
    Thick Gel
    • When pouring a horizontal agarose gel, keep the gel thickness at about 3-4 mm. During electrophoresis, gels with a thickness of more than 5 mm can form in the diffusion strip.
    Didn’t work well
    • Thoroughly clean the gel comb before pouring the gel.
    • To prevent the bottom of the sample from leaking from freezing and smearing of the sample strips, do not push the comb down to the very bottom of the horizontal gel.
    • Avoid overfilling our gel bowls as this can lead to blockage of the wells.
    • Wait for the dents to form and remove the scallop beforehand.
    • Once the gel has hardened, carefully remove the comb and always take care not to damage the impressions.
    invalid type
    • For electrophoresis of bound single-stranded nucleic acids (eg RNA), prepare a final denaturing gel for efficient separation. On the other hand, avoid denaturing gels with double-stranded DNA samples.
    Sample preparation
    Sample not overloaded
    • Use more than really large amounts of samples for fill electrophoresis; Typically 0.1 to 0.2 μg per mm of gel well width is recommended. Stripes, distorted and / or possibly U-shaped stripes, and stripes that appear to merge are a common feature associated with overflowing gels.
    Broken Sample
    • Ensure certain reagents are of molecular biological quality and are typically nuclease-free in laboratory equipment. Follow good laboratory practices (e.g. wearing protective gloves, protect from nuclease contamination, work in specified areas, etc.) when handling acids, nucleic acids, especially when workingwith RNA.
    Sample High Salt Buffers
    • Make sure the salt concentration in the fill buffer is compatible with the selected gel. Thin the loading barrier if necessary.
    • If the nucleic acid pool is already in a high salinity obstruction, dilute the sample to a normal nuclease-free state before adding loading buffer. If necessary, purify or precipitate nucleic acid and resuspend the sample in nuclease-free water to remove more salt.
    High Protein Sample
    • Proteins present in the sample can interfere with the use of sample fluidity in the gel. Remove the protein by purifying the assay, or dissociate / denature the protein by preparing the sample in the SDS load color and heating before loading.
    Incompatible load buffer
    • For electrophoretically linked single-stranded nucleic acids, use a dye-denaturant vehicle, then heat the sample toprevent unwanted duplex formation.
    • For double-stranded DNA electrophoresis, the dye charge is a sufficient reason for denaturation and lack of heating in the experiment to preserve the duplex structure.
    Bubbles on sample loading
    • Make sure air bubbles that appear in the well during Internet sampling are not trapped, avoid tape distortion.
    Well damaged on boot
    • Avoid piercing the pipette tip wells when loading the sample.
    Water sample contains residues containing acrylamide and / or urea
    • If you are using polyacrylamide gels, rinse any remaining acrylamide (and urea if the gels are denatured) from the sample well to load the sample.
    Very low high, also known as voltage
    • Apply the recommended voltage for the specific nucleic acid size range and working buffer used. Very low or high voltage canThis can lead to suboptimal resolution when separating nucleic acids.
    Very short or long controlled time
    • A running gel that lasts long enough for the warranty straps to loosen. However, very long operation can lead to overheating, distorted samples and blurred sounds.
    Incompatible execution buffer
    • Make sure most gels and express buffers are compatible and prepared correctly.
    • Use high-buffered functional electrophoresis for more than 2 hours.
    Sample Render
    Group Money Distribution
    • Avoid storage or better time between completion of electrophoresis and visualization of the gel; smaller molecular size bands can develop as diffuse, the better the nucleic acid staining is documented in the gel.
    Moving lanes
    • Use correct gel tension, commission rate and lead timeTo separate artists with similar molecular sizes. Migratory ribbons often appear as a fuzzy, thick and light collar. Camera
    • To be
    blur Make sure the camera is in focus when frost is visible through the lens for viewing on the screen.

    Why is my gel electrophoresis streaking?

    1. Poorly prepared gel: If the liquid is not poured correctly, it will definitely not polymerize or harden evenly, causing the molecules to smear. If the wells are overfilled, or if the sample has always been poorly diluted, excess melody can smear over the gel.

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