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ALP ELISA Kits
Storage

The plate should be stored at 4oC - 8oC. Do not expose to temperatures in excess of 37oC. Avoid direct sunlight.



List of lab equipments and material required
  • Microcentrifuge
  • Microplate reader
  • Multichannel pipette
  • Deionized (d.i.) water

Criteria for a valid ELISA test/classification of positives
  • Sample wells with all reagents added except seed/leaf extract (blank wells), should have an absorbance value below 0.2 at 405 nm at the time of taking the reading (30-45 min after substrate addition).
  • A positive sample is indicated by a value of 0.2 and above for seed samples and 0.1 for leaf samples, after zeroing the instrument to a blank well.

Sample preparation
[Note: Avoid cross-contamination between samples]
  • For seed extracts: Imbibe cotton seeds overnight in water. Remove seed coat and cut each seed to be tested in half with a clean blade. Place one half of the seed in a microcentrifuge tube and add 500 µl 1X Extraction buffer. Crush with a pestle for 30 seconds. Use 100 µl of this extract for sampling.
    (Refer Recipe will be provided to the customer)
  • For leaf extracts: punch out 3-4 leaf discs with a mcf tube by placing a leaf between the lid and the tube opening and closing the lid onto the leaf. Add 500 µl 1X Extraction buffer, crush with a pestle for 30 seconds. Use 100 µl of this extract for sampling.

Preparation of positive and negative seed extracts

It is important to have Bt positive, Bt negative and blank wells in the test in order to interpret results correctly. Bt positive samples are extracts from established Bt cotton lines. Bt negative samples are extracts from non-Bt plants of the same genotype. Blank wells are those in which no plant extract is added, but otherwise treated identically.

Take 2-4 seeds each of positive and negative lines, separately.

Remove the outer cover of the dry seeds. Crush the seeds to make fine powder, using mortar and pestle. Avoid cross-contamination between positive and negative samples.

Weigh accurately 10 mg each of positive and negative seed sample separately, in a mcf tube. Add 500 µl of 1X Extraction buffer. Crush well with a pestle, allow it to settle. Use 100 µl of this extract per well.

Extracts can be stored frozen at -20o C and re-used for a week.

HRP ELISA Kits
Storage

The plate should be stored at 4o C-8o C. Do not expose to temperatures in excess of 37o C. Avoid direct sunlight.



List of lab equipments and material required

  • Microcentrifuge
  • Microplate reader
  • Multichannel pipette
  • Deionized (d.i.) water


Criteria for a valid ELISA test/classification of positives

Sample wells with all reagents added except seed/leaf extract (blank wells), should have an absorbance value below 0.150 at 450 nm at the time of taking the reading. A positive sample value indicated by a value above than 1 ppb control, after zeroing the instrument to a blank well.



Sample preparation
[Note: Avoid cross-contamination between samples]

  • For seed extracts: Imbibe cotton seeds overnight in water. Remove seed coat and cut each seed to be tested in half with a clean blade. Place one half of the seed in a microcentrifuge tube and add 500 µl 1X Extraction buffer. Crush with a pestle for 30 seconds. Use 100 µl of this extract for sampling.
    (Refer Recipe will be provided to the customer)
  • For leaf extracts: punch out 3-4 leaf discs with a mcf tube by placing a leaf between the lid and the tube opening and closing the lid onto the leaf. Add 500 µl 1X Extraction buffer, crush with a pestle for 30 seconds. Use 100 µl of this extract for sampling.
Xpresstrips™
Xpresstrips™ do not require any specific storing conditions, and can be kept at room temperature during their validity period. For extended shelf life store at 4oC.

Protocols
We offer a range of products as well as protocols, which gives the customer options according to need. Protocols are standardized for highthroughput results:



ELISA kits for qualitative analysis

ALP protocol: (2 Hrs 45 min.)
This ALP based protocol is ideal for testing plants in breeding stages due to its ablilty to distinguish high vs. low expressers.

HRP Protocol: (60 min.)
This HRP based protocol is ideal for qualitative testing of seeds or leaf tissue where upto 10 plates per day are processed. Suitable for testing production lots.



ELISA kits for quantitative analysis

Quantitative Elisa is used for quantification of Bt protein in various plant tissues and can be used for different crops. Quantitative estimation of Bt protein is necessary from the point of gene expression studies and regulatory process. Total time required for one assay is around four hours including the analysis part.



Xpresstrips ™

DesiGen Xpresstrips™ offer rapid and convenient testing of transgenic samples. The high sensitivity of the strips make them ideal for GMO testing. This time saving test gives visual results within 5-10 minutes. Seed or leaf samples can be tested in field without any special training.

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