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The presence of Mg++ ion is required and the optimal concentration is 10mM. T4 DNA ligase has the unique ability to join sticky and blunt ended fragments. Cohesive end ligation is carried out at 12Â°C to 16Â°C to maintain a good balance between annealing of ends and activity of the enzyme.
The test is finished when bleeding has stopped completely. A normal value is less than 9 and a half minutes. A prolonged bleeding time may be a result from decreased number of thrombocytes or impaired blood vessels. However, it should also be noted that the depth of the puncture or incision may be the source of error. lancet or scalpel blade is used to make a shallow incision that is 1 millimeter deep on the underside of the forearm. A standard-sized incision is made around 10 mm long and 1 mm deep. The time from when the incision is made until all bleeding has stopped is measured and is called the bleeding time.